Asgpr-binding compounds for the degradation of extracellular proteins

ABSTRACT

Compounds and compositions that have an asialoglycoprotein receptor (ASGPR) binding ligand bound to an extracellular protein binding ligand for the selective degradation of the target extracellular protein in vivo to treat disorders mediated by the extracellular protein are described.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of International Patent ApplicationNo. PCT/US2021/015939, filed in the U.S. Receiving Office on Jan. 29,2021, which claims the benefit of U.S. Provisional Patent ApplicationNo. 62/968,802, filed Jan. 31, 2020, and U.S. Provisional PatentApplication No. 63/063,015, filed Aug. 7, 2020. The entirety of eachthese applications is incorporated by reference for all purposes.

INCORPORATION BY REFERENCE

The contents of the text file named“19121-001WO1US1_SequenceListing_ST.26.xml” which was created on Jul.28, 2022 and is 78.9 KB in size, are hereby incorporated by reference intheir entirety.

FIELD OF THE INVENTION

This invention provides compounds and compositions that have anasialoglycoprotein receptor (ASGPR) binding ligand bound to anextracellular protein binding ligand for the selective degradation ofthe target extracellular protein in vivo to treat disorders mediated bythe extracellular protein.

BACKGROUND OF THE INVENTION

Historically, therapeutic strategies for the inhibition of proteinsemployed small molecule inhibitors which bound in an enzymatic pocket orat an allosteric position. Those proteins which were not enzymes weredifficult to control, and some were considered “not druggable.”

Intracellular protein degradation is a natural and highly regulated,essential process that maintains cellular homeostasis. The selectiveidentification and removal of damaged, misfolded, or excess proteinswithin the cell is achieved via the ubiquitin-proteasome pathway (UPP).The UPP is central to the regulation of almost all intracellularprocesses. A number of companies and institutions have designedintracellular protein degrading molecules that take advantage of thisnatural process to degrade disease-mediating proteins intracellularly bylinking a ligand to the protein to be degraded to a protein in the UPP.Examples are found in U.S. 2014/0356322 assigned to Yale University,GlaxoSmithKline, and Cambridge EntA122-lerprise Limited University ofCambridge; Buckley et al. (J. Am. Chem. Soc. 2012, 134, 4465-4468)titled “Targeting the Von Hippel-Lindau E3 Ubiquitin Ligase Using SmallMolecules to Disrupt the Vhl/Hif-lalpha Interaction”; WO 2015/160845assigned to Arvinas Inc. titled “Imide Based Modulators of Proteolysisand Associated Methods of Use”; Lu et al. (Chem. Biol. 2015, 22,755-763) titled “Hijacking the E3 Ubiquitin Ligase Cereblon toEfficiently Target Brd4”; Bondeson et al. (Nat. Chem. Biol. 2015, 11,611-617) titled “Catalytic in Vivo Protein Knockdown by Small-MoleculeProtacs”; Gustafson et al. (Angewandte Chemie, International Edition inEnglish 2015, 54, 9659-9662) titled “Small-Molecule-Mediated Degradationof the Androgen Receptor through Hydrophobic Tagging”; Lai et al.(Angewandte Chemie, International Edition in English 2016, 55, 807-810)titled “Modular Protac Design for the Degradation of Oncogenic Bcr-Abl”;Toure et al. (Angew. Chem. Int. Ed. 2016, 55, 1966-1973) titled“Small-Molecule Protacs: New Approaches to Protein Degradation”; Winteret al. (Science 2015, 348, 1376-1381) titled “Drug Development.Phthalimide Conjugation as a Strategy for in Vivo Targeted ProteinDegradation”; U.S. 2016/0058872 assigned to Arvinas, Inc. titled “ImideBased Modulators of Proteolysis and Associated Methods of Use” and U.S.2016/0045607 assigned to Arvinas Inc. titled “Estrogen-related ReceptorAlpha Based PROTAC Compounds and Associated Methods of Use”.

The highjacking of the UPP intracellular process to degrade difficult orundruggable proteins, however, is not available to degrade extracellularproteins. Nonlimiting examples of extracellular proteins includeimmunoglobulins and cytokines, which can play a strong role in creatingor exacerbating serious diseases. Immunoglobulins include IgA, IgG, IgD,IgE, and IgM. Cytokines are cell signaling peptides secreted into thebloodstream which cannot cross the lipid bilayer of cells to enter thecytoplasm, for example, interferons, interleukins, chemokines,lymphokines, MIP, and tumor necrosis factors. Cytokines are involved inautocrine, paracrine and endocrine signaling. They mediate immunity,inflammation and hematopoiesis. Cytokines are produced by immune cells(macrophages, B-cells, T-cells and mast cells), endothelial cells,fibroblasts and stromal cells.

The asialoglycoprotein receptor (ASGPR) is a Ca²⁺—dependent lectin thatis primarily expressed in parenchymal hepatocyte cells. The main role ofASGPRs is to help regulate serum glycoprotein levels by mediatingendocytosis of desialylated glycoproteins (as depicted below). Thereceptor binds ligands with a terminal galactose orN-acetylgalactosamine. The C³— and C⁴-hydroxyl groups bind to Ca²⁺. TheC^(z) N-acetyl position has also been considered important to bindingactivity.

Asialoglycoproteins bind to ASGPRs and are then cleared byreceptor-mediated endocytosis. The receptor and the protein aredissociated in the acidic endosomal compartment and the protein iseventually degraded by lysosomes. The receptor is endocytosed andrecycled constitutively from the endosome back to the plasma membraneabout every 15 minutes regardless of whether or not a glycoprotein isbound. However, it has been shown that the internalization rate of thereceptor is dependent on the presence of ligand. In a 1998 study, theinternalization rate of the protein without ligand was less thanone-third of the rate of internalization of the ligand-receptor complex(Bider et al. FEBS Letters, 1998, 434, 37).

The ASGPR is comprised of two homologous subunits with 58% sequenceidentity known as H1 and H2. Various ratios of H1 and H2 form functionalhomo- and hetero-oligomers with different conformations, but the mostabundant conformation is a trimer composed of two H1 and one H2subunits. The ASGPR is composed of a cytoplasmic domain, a transmembranedomain, a stalk region, and a carbohydrate recognition domain (CRD).Both the H1 and H2 subunit are required to form the CRD, and therefore,co-expression of both subunits is a requirement for endocytosis ofasialoglycoproteins. In 2000, the crystal structure of the CRD regionwas published, revealing three Ca²⁺ binding sites (Meier et al. J. Mol.Biol. 2000, 300, 857).

A number of publications describe ligands that are thought to bind tothe CRD region of ASGPRs. For example, Stokmaier et al. (Bioorg. Med.Chem., 2009, 17, 7254) describes the synthesis of a series of D-GalNAcderivatives where the anomeric OH group is removed and the acetamidogroup is replaced with a 4-substituted 1,2,3-triazole moiety. The mostpotent compound is twice as potent as D-GalNAc in competitive NMRbinding experiments. Mamidyala et al. (JACS, 2012, 134, 1978) describescompounds derived from 2-azidogalactosyl analogs where the anomericposition is occupied by either a β-methyl or a β-4-methoxy-phenyl groupand the azide group is replaced with an amide or a triazole. The ligandswere tested for binding activity by surface plasmon resonance and manyexhibited more potent K_(d) values than that of the parentN-acetylgalactosamine.

Studies have also shown that the receptor affinity for a ligand may beinfluenced by the ligand's valency. For example, Lee et al. (J. Biol.Chem., 1983, 258, 199) showed that the IC₅₀ ranged from approximately 1mM for monoantennary oligosaccharides to approximately 1 nM fortrianternnary oligosaccharides in an assay studying the binding abilityof certain analogs to rabbit hepatocytes.

ASGPRs are primarily expressed on hepatocytes and are minimally found oncells outside of the liver. Hepatocytes exhibit a high exposition ofASGPR binding cites (approximately 100,000-500,000 binding sites percell).

U.S. Pat. No. 5,985,826 to NeoRx Corporation describes the use ofhepatic-directed systems that include a therapeutic agent with activityagainst a liver disease or disorder that is bound to a director moiety.The director moiety, which in one embodiment is a galactose or galactosederivative, directs the active agent to the liver, where the activeagent acts as a therapeutic agent that is then removed from circulationwith assistance from the director moiety.

U.S. Pat. Nos. 9,340,553; 9,617,293; 10,039,778; 10,376,531, and10,813,942 assigned to Pfizer Inc. describe certain bicyclic, bridgedketal derivatives of GalNAc as targeting agents for the ASGPR receptorthat in one embodiment are bound to a linker and/or a therapeutic agentsuch as a small molecule, an amino acid sequence, a nucleic acidsequence, an antibody, or a fluorescent probe. The linker of the drugdelivery system can be monovalent, divalent, or trivalent. Thedisclosure also includes a method for the treatment of a liver diseaseor condition comprising administering the targeted drug delivery system.Several monovalent, divalent, and trivalent bicyclic bridgedGalNAc-derived ASGPR targeting agents linked to fluorescence probes aredisclosed in Sanhueza et al. (JACS, 2017, 139, 3528). One trivalentconjugate in particular exhibited selective hepatocyte targeting in anin vivo biodistribution study in mice.

Pfizer Inc. and the Regents of the University of California jointlydisclosed the use of targeted drug delivery systems comprising certainASPGR targeting ligands covalently bound to a ribonucleoprotein or anendonuclease in US 2017/0137801 for use in CRISPR gene editing.

Pfizer also developed PK2, a targeted drug delivery system whereindoxorubicin is linked via a lysosomally degradable tetrapeptide sequenceto N-(2-hydroxypropyl)methacrylamide copolymers bearing galactosamine asthe targeting agent. In a Phase 1 clinical trial to determine theselectivity, toxicity, and pharmacokinetic profile, it was demonstratedthat the drug targeted primary hepatocellular tumors in patients withprimary or metastatic liver cancer (Seymour et al. J. Clin. Oncol. 2002,20, 1668).

Conjugates of paclitaxel covalently bound to one, two, or three units ofGalNAc via a short linker are described in Petrov et al. (Bioorganic andMedicinal Chemistry Letters, 2018, 28, 382). The analogs were cytotoxicagainst human hepatocellular carcinoma cells and showed high affinityfor ASGPR via surface plasmon resonance.

Pfizer Inc. and Wave Life Sciences Ltd. jointly disclosed the use ofselected ASGPR ligands attached to oligonucleotides in PCT ApplicationsWO 2018/223073 and WO2018/223081. The '073 application describes the useof APOC3 oligonucleotides attached to an ASGPR targeting ligand forselective delivery to the liver and the '081 application describes theuse of PNPLA3 oligonucleotides attached to an ASGPR targeting ligand.PCT Application WO 2018/223056 assigned to Wave Sciences Ltd. describescompositions comprising oligonucleotides for RNA interference and in oneembodiment, the oligonucleotide is attached to an ASGPR targetingligand.

The targeted delivery of antisense oligonucleotides (ASOs), which bindand modulate complementary RNA, to hepatocytes via an ASGPR targetingligand was studied in Schmidt et al. (Nucleic Acids Research, 2017, 45,2294). Mono, di, and trivalent GalNAc were conjugated to single strandedand duplexed ASOs and it was found that di- and trivalentGalNAc-conjugated ASO systems were bind to ASGPR with the strongestaffinity.

Examples of ASGPR-targeted therapy using modified glycoproteins as thetarget agents are reviewed in Huang et al. (Bioconjugate Chem. 2017, 28,283). A number of multivalent ligands that have been developed arediscussed in addition to certain properties for drug delivery, includinglinker length and spatial geometry of the scaffold.

Yale University has filed two PCT Applications, WO 2019/199621 and WO2019/199634, which describe the use of certain ASGPR targeting ligandscovalently bound to a circulating protein binding moiety. Once thecirculating protein binding moiety binds the circulating protein, thecomplex passes to the liver where it is recognized by ASGPR and degradedvia the endo-lysosomal pathway. The '621 application describescirculating protein binding moieties that are capable of targetingmacrophage migration inhibitory factor (MIF) and/or immunoglobulin G(IgG). The '634 application describes the targeting of numerouscirculating proteins including CD40L, TNF-α, PCSK9, VEGF, TGF-β, uPAR,PSMA, IL-2, GP120, TSP-1, and CXCL-2 using a drug delivery systemcomprising a circulating protein binding moiety covalently bound to atargeting ligand, which is a ASGPR targeting ligand.

The Board of Trustees of the Leland Stanford Junior University has fileda PCT application, WO2020/132100, which describes the use of compoundsthat bind a lysosomal targeting molecule such as ASGPR to degrade a cellsurface molecule or extracellular molecule. Compounds related to theWO2020/132100 disclosure are described in an article by Banik et al.(Nature, 2020, 584, 291). Related work from the Bertozzi group waspublished in a preprint article titled “Lysosome Targeting Chimeras(LYTACs) That Engage a Liver-Specific Asialoglycoprotein Receptor forTargeted Protein Degradation,” online on ChemRxiv in July 2020.

While some progress has been made in the area of targeted degradation ofdisease-mediating extracellular proteins, much is left to beaccomplished. There remains an unmet need for additional chemicalcompounds and approaches to treat medical disorders mediated byextracellular proteins.

SUMMARY OF THE INVENTION

Novel compounds and their pharmaceutically acceptable salts andcompositions thereof that degrade disease-mediating extracellularproteins, as well as starting materials and intermediates for suchcompounds and their methods of use and processes of manufacture areprovided. This invention focuses on novel modifications of theC²-position of the ASGPR ligand, referred to herein as R². Thesemodifications include molecules with the C² substituent in the “down”configuration which correspond to the stereochemistry of galactose aswell as molecules with the C² substituent in the “up” configurationwhich corresponds to the stereochemistry of talose. It has beendiscovered that advantageous extracellular protein degrader moleculesare provided when ASGPR ligands with R² groups as specified herein thathave either galactose or talose stereochemistry are incorporated intothe structure.

The extracellular protein degrading compounds described herein can beused to degrade a selected extracellular protein by attaching a ligandfor the extracellular protein to a selected ASGPR ligand, through acovalent bond or a covalent linking group. Extracellular proteins thatcan be targeted according the present invention include but are notlimited to immunoglobulins such as IgA, IgG, IgD, IgE, and IgM, andderivatives thereof which retain the same basic function, and cytokinessuch as interferons, interleukins, chemokines, lymphokines, MIP, andtumor necrosis factors. In certain embodiments, the extracellularprotein is selected from IgA, IgG, IgE, TNF (α or β), IL-1b, IL-2,IFN-γ, IL-6, VGEF, TGF-b1 and PCSK-9. In other nonlimiting embodiments,proteins of the complement system are targeted for degradation,including Factor B, Factor D, Factor H and CC5.

Galactose-Based Molecules

It has been discovered that sugars in the galactose stereochemistry withnew C² substituents are useful ligands for ASGPR. These molecules can beused as ASGPR ligands or linked to an extracellular protein targetingligand to recruit extracellular protein and degrade it in the liver.

In particular, a compound of Formula I, Formula II, Formula III, FormulaIV, Formula V, Formula VI, Formula VII, or Formula VIII is provided:

or a pharmaceutically acceptable salt thereof;

wherein:

X¹ is 1 to 5 contiguous atoms independently selected from O, S, N(R⁶),and C(R⁴)(R⁴), wherein if X¹ is 1 atom then X¹ is O, S, N(R⁶), orC(R⁴)(R⁴), if X¹ is 2 atoms then no more than 1 atom of X¹ is O, S, orN(R⁶), if X¹ is 3, 4, or 5 atoms then no more than 2 atoms of X¹ are O,S, or N(R⁶);

R² is selected from

-   (i) aryl, heterocycle, and heteroaryl containing 1 or 2 heteroatoms    independently selected from N, O, and S, each of which aryl,    heterocycle, and heteroaryl is optionally substituted with 1, 2, 3,    or 4 substituents;

-   (iii) —NR⁸—S(O)—R³, —NR⁸—C(S)—R³, —NR⁸—S(O)(NR⁶)—R³, —N═S(O)(R³)₂,    —NR⁸C(O)NR⁹S(O)₂R³, —NR⁸—S(O)₂—R¹⁰, and —NR⁸—C(NR⁶)—R³ each of which    is optionally substituted with 1, 2, 3, or 4 substituents; and-   (iv) hydrogen, R¹⁰, alkyl-C(O)—R³, —C(O)—R³, alkyl, haloalkyl,    —OC(O)R³, and —NR⁸—C(O)R¹⁰;

R¹⁰ is selected from alkenyl, allyl, alkynyl, —NR⁶-alkenyl, —O-alkenyl,—NR⁶-alkynyl, —NR⁶— heteroaryl, —NR⁶-aryl, —O—heteroaryl, —O—aryl, and—O—alkynyl, each of which R¹⁰ is optionally substituted with 1, 2, 3, or4 substituents;

or R¹⁰ is selected from aryl, alkyl-NR⁸—C(O)—R³, alkyl-aryl,alkyl-heteroaryl with 1, 2, or 4 heteroatoms, alkyl-cyano, alkyl—OR⁶,alkyl—NR⁶R⁸, NR⁸—NR⁶—C(O)R³, NR⁸—S(O)₂—R³, alkenyl, allyl, alkynyl,—NR⁶-alkenyl, —O—alkenyl, —NR⁶-alkynyl, —NR⁶-heteroaryl, —NR⁶-aryl,—O—heteroaryl, —O—aryl, and —O—alkynyl, each of which R¹⁰ is optionallysubstituted with 1, 2, 3, or 4 substituents; in certain embodiments R¹⁰is selected from

in certain embodiments R¹⁰ is selected from

and R⁵ are independently selected from hydrogen, heteroalkyl,C₀-C₆alkyl-cyano, alkyl, alkenyl, alkynyl, haloalkyl, F, Cl, Br, I,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycle,heterocycloalkyl, haloalkoxy, —O—alkenyl, —O—alkynyl, C₀-C₆alkyl-OR⁶,C₀-C₆alkyl—SR⁶, C₀-C₆alkyl—NR⁶R⁷, C₀-C₆alkyl-C(O)R³, C₀-C₆alkyl—S(O)R³,C₀-C₆alkyl-C(S)R³, C₀-C₆alkyl—S(O)₂R³, C₀-C₆alkyl-N(R⁸)—C(O)R³,C₀-C₆alkyl—N(R⁸)—S(O)R³, C₀-C₆alkyl-N(R⁸)—C(S)R³,C₀-C₆alkyl—N(R⁸)—S(O)₂R³ C₀-C₆alkyl—O—C(O)R³, C₀-C₆alkyl—O—S(O)R³,C₀-C₆alkyl—O—C(S)R³, —N═S(O)(R³)₂, C₀-C₆alkylN₃, andC₀-C₆alkyl—O—S(O)₂R³, each of which is optionally substituted with 1, 2,3, or 4 substituents;

R³ at each occurrence is independently selected from hydrogen, alkyl,heteroalkyl, haloalkyl (including —CF₃, —CHF₂, —CH₂F, —CH₂CF₃, —CH₂CH₂F,and —CF₂CF₃), arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl,heteroaryl, heterocycle, —OR⁸, and —NR⁸R⁹;

R⁴ is independently selected at each occurrence from hydrogen,heteroalkyl, alkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl,alkynyl, aryl, heteroaryl, heterocycle, —OR⁶, —NR⁶R⁷, C(O)R³, S(O)R³,C(S)R³, and S(O)₂R³;

R⁶ and R⁷ are independently selected at each occurrence from hydrogen,heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl,haloalkyl, heteroaryl, heterocycle, —alkyl—OR⁸, —alkyl—NR⁸R⁹, C(O)R³,S(O)R³, C(S)R³, and S(O)₂R³;

R⁸ and R⁹ are independently selected at each occurrence from hydrogen,heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl,heteroaryl, and heterocycle;

Cycle is a 3-8 membered fused cyclic group optionally substituted with1, 2, 3, or 4 substituents; exemplary Cycle groups include carbocycle(e.g. cyclopropane, cyclohexane, or cyclohexene), heterocycle (e.g.oxetane, piperazine), aryl (e.g. phenyl), or a heteroaryl group (e.g.pyridine, furan, or pyrrole) as appropriate and allowed by valence;

each Linker^(A) is a bond or a moiety that covalently links the ASGPRligand to Linker^(B);

Linker^(A) is a bond or a moiety that covalently links Linker^(A) to anExtracellular Protein Targeting Ligand;

Extracellular Protein Targeting Ligand is a chemical moiety that bindsto the targeted disease-modifying extracellular protein; and

when a compound is “optionally substituted” it may be substituted asallowed by valence by one or more groups selected from alkyl (includingC₁-C₄alkyl), alkenyl (including C₂-C₄alkenyl), alkynyl (includingC₂-C₄alkynyl), haloalkyl (including C₁-C₄haloalkyl), —OR⁶, F, Cl, I, Br,—NR⁶R⁷, heteroalkyl, cyano, nitro, C(O)R³

wherein the optional substituent is selected such that a stable compoundresults.

In an alternative embodiment when a compounds is “optionallysubstituted” it may be substituted as allowed by valence with one ormore groups selected from alkyl (including C₁-C₄alkyl), alkenyl(including C₂-C₄alkenyl), alkynyl (including C₂-C₄alkynyl), haloalkyl(including C₁-C₄haloalkyl), —OR⁶, F, Cl, Br, I, —NR⁶R⁷, heteroalkyl,heterocycle, heteroaryl, aryl, cyano, nitro, hydroxyl, azide, amide,—SR³, —S(O)(NR⁶)R³, —NR⁸C(O)R³, —C(O)NR⁶R⁷, —C(O)OR³, —C(O)R³, —SF₅,

wherein the optional substituent is selected such that a stable compoundresults.

In one embodiment the Extracellular Protein Targeting Ligand is not anoligomer.

In another embodiment neither the Extracellular Protein nor theExtracellular Protein Targeting Ligand directly mediates intracellulargene editing such as CRISPR.

In an alternative embodiment of the invention, when R² is NR⁶-alkenyl,—NR⁶-alkynyl,—NR⁸-C(O)R¹⁰, —NR⁸—S(O)₂-alkenyl, —NR⁸—S(O)₂-alkynyl,—NR⁶-heteroaryl, or —NR⁶-aryl, then Extracellular Protein TargetingLigand does not comprise an oligonucleotide. In certain embodiment ofthe invention, when R² is R¹⁰, NR⁶-alkenyl, —NR⁶-alkynyl,—NR⁸—C(O)R¹⁰,—NR⁸—S(O)₂-alkenyl, —NR⁸—S(O)₂-alkynyl, —NR⁶-heteroaryl, or —NR⁶-aryl,then Extracellular Protein Targeting Ligand does not comprise anoligonucleotide.

A compound of Formula I-Bi, Formula II-Bi, Formula III-Bi, FormulaIV-Bi, Formula V-Bi, Formula VI-Bi, Formula VII-Bi, or Formula VIII-Biis provided:

or a pharmaceutically acceptable salt thereof;wherein:

Linker^(C) is a chemical group that links each Linker^(A) to theExtracellular Protein Targeting Ligand; and

all other variables are as defined herein.

A compound of Formula I-Tri, Formula II-Tri, Formula III-Tri, FormulaIV-Tri, Formula V-Tri, Formula VI-Tri, Formula VII-Tri, or FormulaVIII-Tri is also provided:

or a pharmaceutically acceptable salt thereof;wherein:

Linker^(D) is a chemical group that links each Linker^(A) to theExtracellular Protein Targeting Ligand; and

all other variables are as defined herein.

As used herein, Anchor Bond is defined as the chemical bond between theExtracellular Protein Targeting Ligand and either Linker^(B), Linker^(C)or Linker^(D), as appropriate.

A compound of Formula IX, Formula X, Formula XI, Formula XII, or FormulaXIII is provided:

or a pharmaceutically acceptable salt thereof;

wherein:

R^(L) is selected from R⁵ and Linker^(E);

R^(L2) is selected from R⁶ and Linker^(E);

X¹ is 1 to 5 contiguous atoms independently selected from O, S, N(R⁶),and C(R⁴)(R⁴), wherein if X¹ is 1 atom then X¹ is O, S, N(R⁶), orC(R⁴)(R⁴), if X¹ is 2 atoms then no more than 1 atom of X¹ is O, S, orN(R⁶), if X¹ is 3, 4, or 5 atoms then no more than 2 atoms of X¹ are O,S, or N(R⁶);

R^(2A) is selected from

-   (i) aryl, heterocycle, and heteroaryl containing 1 or 2 heteroatoms    independently selected from N, O, and S, each of which aryl,    heterocycle, and heteroaryl is optionally substituted with 1, 2, 3,    or 4 substituents;

-   (iii) —NH—S(O)—R³, —NR⁸—C(S)—R³, —NH—S(O)(NR⁶)—R³, and    —N═S(O)(R³)—NR⁶R⁷ each of which is optionally substituted with 1, 2,    3, or 4 substituents;

In an alternative embodiment R^(2A) is selected from

In an alternative embodiment R^(2A) is selected from R¹⁰;

R¹ and R⁵ are independently selected from hydrogen, heteroalkyl,C₀-C₆alkyl-cyano, alkyl, alkenyl, alkynyl, haloalkyl, F, Cl, Br, I,aryl, arylalkyl, heteroaryl, heteroarylalkyl, heterocycle,heterocycloalkyl, haloalkoxy, —O—alkenyl, —O—alkynyl, C₀-C₆alkyl-OR⁶,C₀-C₆alkyl—SR⁶, C₀-C₆alkyl—NR⁶R⁷, C₀-C₆alkyl-C(O)R³, C₀-C₆alkyl—S(O)R³,C₀-C₆alkyl-C(S)R³, C₀-C₆alkyl—S(O)₂R³, C₀-C₆alkyl—N(R⁸)—C(O)R³,C₀-C₆alkyl—N(R⁸)—S(O)R³, C₀-C₆alkyl-N(R⁸)—C(S)R³,C₀-C₆alkyl—N(R⁸)—S(O)₂R³ C₀-C₆alkyl—O—C(O)R³, C₀-C₆alkyl—O—S(O)R³,C₀-C₆alkyl—O—C(S)R³, —N═S(O)(R³)₂, C₀-C₆alkylN₃, andC₀-C₆alkyl—O—S(O)₂R³, each of which is optionally substituted with 1, 2,3, or 4 substituents;

R³ at each occurrence is independently selected from hydrogen, alkyl,heteroalkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl,aryl, heteroaryl, heterocycle, —OR⁸, or —NR⁸R⁹

R⁴ is independently selected at each occurrence from hydrogen,heteroalkyl, alkyl, haloalkyl, arylalkyl, heteroarylalkyl, alkenyl,alkynyl, aryl, heteroaryl, heterocycle, —OR⁶, —NR⁶R⁷, C(O)R³, S(O)R³,C(S)R³, and S(O)₂R³;

R⁶ and R⁷ are independently selected at each occurrence from hydrogen,heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl,haloalkyl, heteroaryl, heterocycle, —alkyl—OR⁸, -alkyl—NR⁸R⁹, C(O)R³,S(O)R³, C(S)R³, and S(O)₂R³;

R⁸ and R⁹ are independently selected at each occurrence from hydrogen,heteroalkyl, alkyl, arylalkyl, heteroarylalkyl, alkenyl, alkynyl, aryl,heteroaryl, and heterocycle;

Cycle is a 3-8 membered fused cyclic group optionally substituted with1, 2, 3, or 4 substituents; exemplary Cycle groups include carbocycle(e.g. cyclopropane, cyclohexane, or cyclohexene), heterocycle (e.g.oxetane, of piperazine), aryl (e.g. phenyl), or a heteroaryl group (e.g.pyridine, furan, or pyrrole) as appropriate and allowed by valence;Linker^(E) is

R³⁰ is selected from C₁, Br, I, —NR⁶H, —OH, —N₃, —SH,

—C(O)N(CH₃)OCH₃, —B(OR⁶)(OR⁷), heterocycle, —NR⁶COR³, —OCOR³ and —COR³,

R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹”, and R¹⁹ are independently ateach occurrence selected from the group consisting of a bond, alkyl,—C(O)—, —C(O)O—, —OC(O)—, —SO₂—, —S(O)—, —C(S)—, —C(O)NR⁶—, —NR⁶C(O)—,—O—, —S—, —NR—, —C(R²¹R²¹)—, —P(O)(OR⁶)O—, —P(O)(OR⁶)—,—P(O)(NR⁶R⁷)NR⁶—, —P(O)(NR⁶R⁷)—, amino acid, alkenyl, alkynyl,haloalkyl, alkoxy, aryl, heterocycle, heteroaryl, —[—(CH₂)₂—O—]]—,—[O—(CH₂)₂]_(n)—, —[O—CH(CH₃)C(O)]_(n)—, —[C(O)—CH(CH₃)—O]_(n)—,—[O—CH₂C(O)]_(n)—, —[C(O)—CH₂—O]_(n)—, fatty acid, unsaturated acid,each of which is optionally substituted with 1, 2, 3, or 4 substituentsindependently selected from R²¹;

n is independently selected at each instance from 0, 1, 2, 3, 4, 5, 6,7, 8, 9, or 10;

R²¹ is independently at each occurrence selected from the groupconsisting of hydrogen, alkyl, alkenyl, alkynyl, F, Cl, Br, I, hydroxyl,alkoxy, azide, amino, cyano, —NR⁶R⁷, —NR⁸SO₂R³, —NR'S(O)R³, haloalkyl,heteroalkyl, aryl, heteroaryl, and heterocycle; or

or R²¹ is independently at each occurrence selected from the groupconsisting of hydrogen, alkyl, alkenyl, alkynyl, F, Cl, Br, I, hydroxyl,alkoxy, azide, amino, cyano, —NR⁶R⁷, —NR⁸SO₂R³, —NR⁸S(O)R³, haloalkyl,heteroalkyl, aryl, heteroaryl, heterocyclyl, —SR³, —C(O)OR³,—C(O)NR⁶NR⁸, —OR³,

and heterocycle.

Talose-Based Molecules

It has also been discovered that sugars in the talose stereochemistrywith specific C² substituents are useful ligands for ASGPR. Thesemolecules can be used as ASGPR ligands or linked to an extracellularprotein targeting ligand to recruit extracellular protein and degrade itin the liver.

In particular, a compound of Formula I-d, Formula II-d, Formula III-d,Formula IV-d, Formula V-d, or Formula VI-d is provided:

or a pharmaceutically acceptable salt thereof;

wherein for compounds of Formula I-d, II-d, III-d, IV-d, V-d, and VI-dR² is selected from:

-   (i) aryl, heterocycle, and heteroaryl containing 1 or 2 heteroatoms    independently selected from N, O, and S, each of which aryl,    heterocycle, and heteroaryl is optionally substituted with 1, 2, 3,    or 4 substituents;

-   (iii) —NR⁸—S(O)—R³, —NR⁸—C(S)—R³, —NR⁸—S(O)(NR⁶)—R³, —N═S(O)(R³)₂,    —NR⁸C(O)NR⁹S(O)₂R³, —NR⁸—S(O)₂—R¹⁰, and —NR⁸—C(NR⁶)—R³ each of which    is optionally substituted with 1, 2, 3, or 4 substituents; and-   (iv) hydrogen, R¹⁰, alkyl—C(O)—R³, —C(O)—R³, alkyl, haloalkyl,    —OC(O)R³, and —NR⁸—C(O)R¹⁰; and-   (v) R²⁰⁰; R²⁰⁰ is —NR⁸—C(O)—R³;

or R²⁰⁰ is —NR⁸—C(O)—R³, —NR⁶-alkyl, —OR⁸, heteroaryl (including forexample triazole and tetrazole), NR⁸—S(O)₂—R³, or —NR⁶-heteroalkyl, eachof which R²⁰⁰ substituents is optionally substituted with 1, 2, 3, or 4substituents;

when compounds are “optionally substituted” they may be substituted asallowed by valence by groups selected from alkyl (including C₁-C₄alkyl),alkenyl (including C₂-C₄alkenyl), alkynyl (including C₂-C₄alkynyl),haloalkyl (including C₁-C₄haloalkyl), —OR⁶, F, Cl, Br, I, —NR⁶R⁷,heteroalkyl, cyano, nitro, C(O)R³,

wherein the optional substituent is selected such that a stable compoundresults.

In an alternative embodiment when compounds are “optionally substituted”they may be substituted as allowed by valence by groups selected fromalkyl (including C₁-C₄alkyl), alkenyl (including C₂-C₄alkenyl), alkynyl(including C₂-C₄alkynyl), haloalkyl (including C₁-C₄haloalkyl), —OR⁶, F,Cl, Br, I, —NR⁶R⁷, heteroalkyl, heterocycle, heteroaryl, aryl, cyano,nitro, hydroxyl, azide, amide, —SR³, —S(O)(NR⁶)R³, —NR⁸C(O)R³,—C(O)NR⁶R⁷, —C(O)OR³, —C(O)R³, —SF₅,

wherein the optional substituent is selected such that a stable compoundresults; and

all other variables are as defined herein.

In certain embodiments, a mixture of the galactose and talose-basedstereochemistry are used in medical therapy, including but not limitedto an equal mixture. For example, a compound Formula I and acorresponding compound of Formula I-d may be used in any mixture thatprovides the desired therapeutic results. More generally, any mixture ofany of the Formulas I through XVI and Formulas I-d through XVI-d (any ofwhich can be in the mono, bi, or tri framework).

A compound of Formula I-d-Bi, Formula II-d-Bi, Formula III-d-Bi, FormulaIV-d-Bi, Formula V-d-Bi, or Formula VI-d-Bi is provided:

wherein for compounds of Formula I-d-Bi, II-d-Bi, III-d-Bi, IV-d-Bi,V-d-Bi, and VI-d-Bi, R² is selected from:

-   (i) aryl, heterocycle, and heteroaryl containing 1 or 2 heteroatoms    independently selected from N, O, and S, each of which aryl,    heterocycle, and heteroaryl is optionally substituted with 1, 2, 3,    or 4 substituents;

-   (iii) —NR⁸—S(O)—R³, —NR⁸—C(S)—R³, —NR⁸—S(O)(NR⁶)—R³, —N═S(O)(R³)₂,    —NR⁸C(O)NR⁹S(O)₂R³, —NR⁸—S(O)₂—R¹⁰, and —NR⁸—C(NR⁶)—R³ each of which    is optionally substituted with 1, 2, 3, or 4 substituents; and-   (iv) hydrogen, R¹⁰, alkyl—C(O)—R³, —C(O)—R³, alkyl, haloalkyl,    —OC(O)R³, and —NR⁸—C(O)R¹⁰; and-   (v) R²⁰⁰; R²⁰⁰ is —NR⁸—C(O)—R³; and

all other variables are as defined herein.

A compound of Formula I-d-Tri, Formula II-d-Tri, Formula III-d-Tri,Formula IV—d-Tri, Formula V-d-Tri, or Formula VI-d-Tri, is provided:

or a pharmaceutically acceptable salt thereof, wherein for compounds ofFormula I-d-Tri, II-d-Tri, III-d-Tri, IV-d-Tri, V-d-Tri, and VI-d-Tri R²is selected from:

-   (i) aryl, heterocycle, and heteroaryl containing 1 or 2 heteroatoms    independently selected from N, O, and S, each of which aryl,    heterocycle, and heteroaryl is optionally substituted with 1, 2, 3,    or 4 substituents;

-   (iii) —NR⁸—S(O)—R³, —NR⁸—C(S)—R³, —NR⁸—S(O)(NR⁶)—R³, —N═S(O)(R³)₂,    —NR⁸C(O)NR⁹S(O)₂R³, —NR⁸—S(O)₂—R¹⁰, and —NR⁸—C(NR⁶)—R³ each of which    is optionally substituted with 1, 2, 3, or 4 substituents; and-   (iv) hydrogen, R¹⁰, alkyl—C(O)—R³, —C(O)—R³, alkyl, haloalkyl,    —OC(O)R³, and —NR⁸—C(O)R¹⁰; and-   (v) R²⁰⁰;

R²⁰⁰ is —NR⁸—C(O)—R³; and

all other variables are as defined herein.

A compound of Formula IX-d, X-d, XI-d, XII-d, XIII-d, XIV-d is provided.

or a pharmaceutically acceptable salt thereof.

In an embodiment of the invention, the Extracellular Protein TargetingLigand is a small organic molecule (i.e., a non-biologic) thatadequately binds to the protein in such a manner that it is able totransport it to the liver, the residue of a pharmaceutically activecompound that binds to the target extracellular protein (for example butnot limited to a compound of the sort that would be reviewed as a drugby CDER of the FDA, or an approved or clinical stage drug) or a peptide,protein or biologic or a binding fragment thereof that adequately bindsto the protein in such a manner that it is able to transport it to theliver, and in some embodiments, that does not comprise anoligonucleotide or aptamer. A plethora of illustrative nonlimitingexamples of extracellular protein targeting ligands is provided in FIG.1 . The present invention focuses on the degradation of circulatingextracellular proteins that mediate diseases, for example, involvingimmunity, inflammation, hematopoiesis/blood disorders (including thosecaused or exacerbated by blood vessel formation) and abnormal cellularproliferation such as tumors and cancer. In a typical embodiment of theinvention, neither the Extracellular Protein nor the ExtracellularProtein Targeting Ligand directly mediates intracellular gene editingsuch as CRISPR.

In one embodiment of the invention, when R^(z) is NR⁶-alkenyl,—NR⁶-alkynyl,—NR⁸—C(O)R¹⁰, —NR⁸—S(O)₂-alkenyl, —NR⁸—S(O)₂-alkynyl,—NR⁶-heteroaryl, or —NR⁶-aryl, then Extracellular Protein TargetingLigand does not comprise an oligonucleotide or aptamer.

The ASGPR-binding Extracellular Protein degraders of the presentinvention can be administered in any manner that allows the degrader tobind to the Extracellular Protein, typically in the blood stream, andcarry it to the ASGPR-bearing hepatocyte cells on the liver forendocytosis and degradation. As such, examples of methods to deliver thedegraders of the present invention include, but are not limited to,oral, intravenous, buccal, sublingual, subcutaneous and transnasal.

BRIEF DESCRIPTION OF FIGURES

FIG. 1A provides a non-limiting list of Extracellular Protein TargetingLigands that target Immunoglobulin A (IgA).

FIG. 1B provides a non-limiting list of Extracellular Protein TargetingLigands that target Immunoglobulin G (IgG).

FIG. 1C-1G provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Immunoglobulin E (IgE).

FIG. 1H-1M provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Tumor Necrosis Factor alpha (TNF-α).

FIG. 1N provides a non-limiting list of Extracellular Protein TargetingLigands that target Interleukin-1 (IL-1).

FIG. 1O-1S provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Interleukin-2 (IL-2).

FIG. 1T-1W provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Interleukin-6 (IL-6).

FIG. 1X-IAA provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Interferon gamma (IFN-γ).

FIG. 1BB-1KK provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Vascular endothelial growth factor (VEGF).

FIG. 1LL provides a non-limiting list of Extracellular Protein TargetingLigands that target Transforming growth factor beta (TGF-β1).

FIG. 1MM-1PP provides a non-limiting list of Extracellular ProteinTargeting Ligands that target proprotein convertase subtilisin kexin 9(PCSK-9).

FIG. 1QQ-1SS provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Carboxypeptidase B2 (CPB2).

FIG. 1TT-1UU provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Cholinesterase (ChE).

FIG. 1VV-1WW provides a non-limiting list of Extracellular ProteinTargeting Ligands that target C-C Motif Chemokine Ligand 2 (CCL2).

FIG. 1XX-1BBB provides a non-limiting list of Extracellular ProteinTargeting Ligands that target coagulation factor VII (Factor VII).

FIG. 1CCC-1FFF provides a non-limiting list of Extracellular ProteinTargeting Ligands that target coagulation factor IX (Factor IX).

FIG. 1GGG provides a non-limiting list of Extracellular ProteinTargeting Ligands that target CD40 Ligand (CD40L).

FIG. 1HHH-1JJJ provides a non-limiting list of Extracellular ProteinTargeting Ligands that target coagulation factor Xa (Factor Xa).

FIG. 1KKK-1MMM provides a non-limiting list of Extracellular ProteinTargeting Ligands that target coagulation factor XI (Factor XI).

FIG. 1NNN and 1OOO provides a non-limiting list of Extracellular ProteinTargeting Ligands that target coagulation factor XII (Factor XII).

FIG. 1PPP and 1QQQ provides a non-limiting list of Extracellular ProteinTargeting Ligands that target coagulation factor XIII (Factor XIII).

FIG. 1RRR-1UUU provides a non-limiting list of Extracellular ProteinTargeting Ligands that target fibroblast growth factor 1 (FGF1).

FIG. 1VVV-1XXX provides a non-limiting list of Extracellular ProteinTargeting Ligands that target fibroblast growth factor 2 (FGF2).

FIG. 1YYY and 1ZZZ provides a non-limiting list of Extracellular ProteinTargeting Ligands that target fibronectin (FN1).

FIG. 1AAAA and 1BBBB provides a non-limiting list of ExtracellularProtein Targeting Ligands that target Interleukin-5 (IL-5).

FIG. 1CCCC provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Interleukin-8 (IL-8).

FIG. 1DDDD and 1EEEE provides a non-limiting list of ExtracellularProtein Targeting Ligands that target Interleukin-10 (IL-10).

FIG. 1FFFF and 1GGGG provides a non-limiting list of ExtracellularProtein Targeting Ligands that target Interleukin-21 (IL-21).

FIG. 1HIUH and 1III provides a non-limiting list of ExtracellularProtein Targeting Ligands that target Interleukin-22 (IL-22).

FIG. 1JJJJ-1NNNN provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Kallikrein 1.

FIG. 1OOOO provides a non-limiting list of Extracellular ProteinTargeting Ligands that target lipoprotein lipase (LPL).

FIG. 1PPPP and 1QQQQ provides a non-limiting list of ExtracellularProtein Targeting Ligands that target matrix metalloproteinase-1 (MMP1).

FIG. 1RRRR-1DDDDD provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Macrophage migration inhibitory factor(MIF), also known as glycosylation-inhibiting factor (GIF), L-dopachromeisomerase, or phenylpyruvate tautomerase.

FIG. 1EEEEE-1GGGGG provides a non-limiting list of Extracellular ProteinTargeting Ligands that target neutrophil elastase (NE).

FIG. 1HHHHH and 1IIIII provides a non-limiting list of ExtracellularProtein Targeting Ligands that target Prothrombin.

FIG. 1JJJJJ-1NNNNN provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Plasma kallikrein (KLKB1).

FIG. 1OOOOO-1SSSSS provides a non-limiting list of Extracellular ProteinTargeting Ligands that target plasminogen (PLG).

FIG. 1TTTTT-1XXXXX provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Plasminogen activator inhibitor-1 (PAI-1),endothelial plasminogen activator inhibitor or serpin El.

FIG. 1YYYYY-IAAAAAA provides a non-limiting list of ExtracellularProtein Targeting Ligands that target phospholipases A2, for exampletype 1B or group 1B (PLA2, PA21B, PLA2G1B, PLA2-IB).

FIG. 1BBBBBB-1DDDDDD provides a non-limiting list of ExtracellularProtein Targeting Ligands that target phospholipases A2, for exampletype IIA or group IIA (PLA2, PLA2A, PA2IIA, PLA2G2A, PLA2-IIA).

FIG. 1EEEEEE-1NNNNNN provides a non-limiting list of ExtracellularProtein Targeting Ligands that target placental growth factor (PGF).

FIG. 1OOOOOO-1QQQQQQ provides a non-limiting list of ExtracellularProtein Targeting Ligands that target plasminogen activator, tissue type(tPA, PLAT).

FIG. 1RRRRRR provides a non-limiting list of Extracellular ProteinTargeting Ligands that target Transforming growth factor beta 2 (TGF-β2,TGFB2).

FIG. 1SSSSSS provides a non-limiting list of Extracellular ProteinTargeting Ligands that target thrombospondin 1 (TSP1, TSP-1, THBS1).

FIG. 1TTTTTT-1XXXXXX provides a non-limiting list of ExtracellularProtein Targeting Ligands that target Urokinase or Urokinase-typeplasminogen activator (UPA, uPA).

FIG. 2 provides a non-limiting list of exemplary Extracellular ProteinTargeting Ligands that target complement factor B.

FIG. 3A and 3B provides a non-limiting list of exemplary ExtracellularProtein Targeting Ligands that target complement factor D.

FIG. 4 provides a non-limiting list of exemplary Extracellular ProteinTargeting Ligands that target complement factor H.

FIG. 5 provides a non-limiting list of exemplary Extracellular ProteinTargeting Ligands that target complement component 5.

FIG. 6 provides a non-limiting list of exemplary Extracellular ProteinTargeting Ligands that target TNF-alpha.

FIG. 7 provides a non-limiting list of exemplary Extracellular ProteinTargeting Ligands that target factor XI.

FIG. 8 provides a non-limiting list of exemplary formulas of the presentinvention.

DETAILED DESCRIPTION OF THE INVENTION

Novel compounds and their pharmaceutically acceptable salts andcompositions thereof that degrade disease-mediating extracellularproteins, as well as starting materials and intermediates for suchcompounds and their methods of use and processes of manufacture areprovided. This invention focuses on novel modifications of theC²-position of the ASGPR ligand, referred to herein as R². Thesemodifications include molecules with the C² substituent in the “down”configuration which correspond to the stereochemistry of galactose aswell as molecules with the C² substituent in the “up” configurationwhich corresponds to the stereochemistry of talose. It has beendiscovered that advantageous extracellular protein degrader moleculesare provided when ASGPR ligands with R² groups as specified herein thathave either galactose or talose stereochemistry are incorporated intothe structure.

I. Galactose-Based ASGPR-Binding Extracellular Protein Degraders of thePresent Invention

As used in the embodiments here, xx is independently selected from 0, 1,2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19,20,21,22,23,24,and 25.

As used in the embodiments here, yy is independently selected from 0, 1,2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19,20,21,22,23,24,and 25.

As used in the embodiments here, zz is independently selected from 0, 1,2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19,20,21,22,23,24,and 25.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound ofthe present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound o the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In one aspect of the present invention an Extracellular Proteindegrading compound is provided wherein the ASGPR ligand is aliganddescribed herein

in this aspect the ASGPR ligand is linked in either the C1 or C5 (R¹ orR⁵) position to form a degrading compound, for example, when the ASGPRligand is

then non-limiting examples of ASGPR binding compounds contemplated bythis aspect include:

or the bi- or tri-version thereof or a pharmaceutically acceptable saltthereof.

In any of the embodiments herein where an ASGPR ligand is drawn for usein a degrader the ASGPR ligand is typically linked through to theExtracellular Protein Targeting Ligand in the C⁵ position (e.g., whichcan refer to the adjacent C⁶ carbon hydroxyl or other functional moietythat can be used for linking purposes). When the linker andExtracellular Protein Targeting Ligand is connected through the C¹position, then that carbon is appropriately functionalized for linking,for example with a hydroxyl, amino, allyl, alkyne or hydroxyl-allylgroup. Typically the ASGPR ligand is not linked in the C³ or C⁴position, because these positions chelate with the calcium for ASGPRbinding in the liver.

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention is

In certain embodiments, the compound of the present invention is

II. Talose-Based ASGPR-Binding Extracellular Protein Degraders of thePresent Invention

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compoundof thepresent invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom.

In certain embodiments the compound of the present invention is selectedfrom:

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In certain embodiments the compound of the present invention is selectedfrom:

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In certain embodiments the compound of the present invention is selectedfrom:

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In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

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In certain embodiments the compound of the present invention is selectedfrom:

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In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom.

In certain embodiments the compound of the present invention is selectedfrom.

In certain embodiments the compound of the present invention is selectedfrom

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In one aspect of the present invention an Extracellular Proteindegrading compound is provided wherein the ASGPR ligand is a ligand asdescribed herein

in this aspect the ASGPR ligand is linked in either the C1 or C5 (R¹ orR⁵) position to form a degrading compound, for example, when the ASGPRligand is

then non-limiting examples of ASGPR binding compounds contemplated bythis aspect include:

or the bi- or tri-version thereof or a pharmaceutically acceptable saltthereof.

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR³,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

wherein in certain embodiments R² is selected from —NR⁶COR¹⁰,—NR⁶-(5-membered heteroaryl), and—NR⁶-(6-membered heteroaryl), each ofwhich R² groups is optionally substituted with 1, 2, 3, or 4 independentsubstituents as described herein, for example 1, 2, 3, or 4substitutents independently selected from F, Cl, Br, haloalkyl, oralkyl.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom.

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom:

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, an ASGPR ligand useful for incorporation into acompound of the present invention is selected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

In certain embodiments, the compound of the present invention isselected from

III. Embodiments of the ASGPR Ligand Embodiments of R¹

In certain embodiments R¹ is hydrogen.

In certain embodiments R¹ is

In certain embodiments R¹ is

In certain embodiments R¹ is

In certain embodiments R¹ is

In certain embodiments R¹ is

In certain embodiments R is

In certain embodiments R¹ is heteroalkyl optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R¹ is C₀-C₆alkyl-cyano optionally substitutedwith 1, 2, 3, or 4 substituents.

In certain embodiments R¹ is alkyl optionally substituted with 1, 2, 3,or 4 substituents.

In certain embodiments R¹ is alkenyl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R¹ is alkynyl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R¹ is haloalkyl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R¹ is F.

In certain embodiments R¹ is Cl.

In certain embodiments R¹ is Br.

In certain embodiments R¹ is aryl optionally substituted with 1, 2, 3,or 4 substituents.

In certain embodiments R¹ is arylalkyl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R¹ is heteroaryl optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R¹ is heteroarylalkyl optionally substituted with1, 2, 3, or 4 substituents.

In certain embodiments R¹ is heterocycle optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R¹ is heterocycloalkyl optionally substitutedwith 1, 2, 3, or 4 substituents.

In certain embodiments R¹ is haloalkoxy optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R¹ is —O—alkenyl, —O—alkynyl, C₀-C₆alkyl—OR⁶,C₀-C₆alkyl—SR⁶, C₀-C₆alkyl—NR⁶R⁷, C₀-C₆alkyl—C(O)R³, C₀-C₆alkyl—S(O)R³,C₀-C₆alkyl-C(S)R³, C₀-C₆alkyl—S(O)₂R³, C₀-C₆alkyl—N(R⁸)—C(O)R³,C₀-C₆alkyl—N(R⁸)—S(O)R³, C₀-C₆alkyl—N(R⁸)—C(S)R³,C₀-C₆alkyl—N(R⁸)—S(O)₂R³ C₀-C₆alkyl—O—C(O)R³, C₀-C₆alkyl—O—S(O)R³,C₀-C₆alkyl—O—C(S)R³, —N═S(O)(R³)₂, C₀-C₆alkylN₃, orC₀-C₆alkyl—O—S(O)₂R³, each of which is optionally substituted with 1, 2,3, or 4 substituents.

Embodiments of R²

In certain embodiments R² is aryl optionally substituted with 1, 2, 3,or 4 substituents.

In certain embodiments R² is heterocycle optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is heteroaryl containing 1 or 2 heteroatomsindependently selected from N, O, and S optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is selected from

In certain embodiments R² is heterocycle optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —NR⁸—S(O)—R³ optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —NR⁸—C(S)—R³ optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —NR⁸—S(O)(NR⁶)—R³ optionally substitutedwith 1, 2, 3, or 4 substituents.

In certain embodiments R² is —N═S(O)(R³)₂ optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —NR⁸C(O)NR⁹S(O)₂R³ optionally substitutedwith 1, 2, 3, or 4 substituents.

In certain embodiments R² is —NR⁸—S(O)₂—R¹⁰ optionally substituted with1, 2, 3, or 4 substituents.

In certain embodiments R² is —NR⁸—C(NR⁶)—R³ optionally substituted with1, 2, 3, or 4 substituents.

In certain embodiments R² is hydrogen.

In certain embodiments R² is R¹⁰.

In certain embodiments R² is alkyl—C(O)—R³.

In certain embodiments R² is —C(O)—R³.

In certain embodiments R² is alkyl.

In certain embodiments R² is haloalkyl.

In certain embodiments R² is —OC(O)R³.

In certain embodiments R² is —NR⁸—C(O)R¹⁰.

In certain embodiments R² is alkenyl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R² is allyl optionally substituted with 1, 2, 3,or 4 substituents.

In certain embodiments R² is alkynyl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R² is —NR⁶-alkenyl optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —O—alkenyl optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —NR⁶-alkynyl optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is —NR⁶-heteroaryl optionally substituted with1, 2, 3, or 4 substituents.

In certain embodiments R² is —NR⁶-aryl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R² is —O—heteroaryl optionally substituted with1, 2, 3, or 4 substituents.

In certain embodiments R² is —O—aryl optionally substituted with 1, 2,3, or 4 substituents.

In certain embodiments R² is —O—alkynyl optionally substituted with 1,2, 3, or 4 substituents.

In certain embodiments R² is selected from

In certain embodiments R² is selected from:

In certain embodiments R² is selected from

wherein R is an optional substituent as defined herein.

In certain embodiments R² is selected from

In certain embodiments R^(2A) is selected from

wherein R is an optional substituent as defined herein.

In certain embodiments R^(2A) is selected from

In certain embodiments, R² is selected from

In certain embodiments, R² 13 selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R⁸ is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is or R^(2A) selected from

In certain embodiments R² is selected from

In certain embodiments R² is selected from

In certain embodiments R² is

In certain embodiments R² is

In certain embodiments R² is a spirocyclic heterocycle for example

In certain embodiments R² is a silicon containing heterocycle forexample

In certain embodiments R² is substituted with SF₅ for example

In certain embodiments R² is substituted with a sulfoxime for example

Embodiments of R¹⁰

In certain embodiments, R¹⁰ is selected from bicyclic heterocycle.

In certain embodiments, R¹⁰ is selected from spirocyclic heterocycle.

In certain embodiments, R¹⁰ is selected from —NR⁶-heterocycle.

In certain embodiments, R¹⁰ is selected from

In certain embodiments, R¹⁰ is selected from

In certain embodiments, R¹⁰ is selected from

In certain embodiments, R¹⁰ is selected from

Embodiments of Cycle

In certain embodiments Cycle is selected from

Embodiments of R³⁰

In one embodiment R³⁰ is selected from:

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

In certain embodiments R²⁰⁰ is

IV. Embodiments of the Linker

In non-limiting embodiments, Linker^(A) and Linker^(B) are independentlyselected from: wherein:

R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R⁸, R¹⁹, and R²⁰ are independently ateach occurrence selected from the group consisting of a bond, alkyl,—C(O)—, —C(O)O—, —OC(O)—, —S02—, —S(O)—, —C(S)—, —C(O)NR⁶—, —NR⁶C(O)—,—O—, —S—, —NR⁶—, —C(R²¹R²¹)—, —P(O)(R³)₀—, —P(O)(R³)—, a divalentresidue of a natural or unnatural amino acid, alkenyl, alkynyl,haloalkyl, alkoxy, aryl, heterocycle, heteroaryl,—CH₂CH₂—[O—(CH₂)₂]_(n)—O—, —CH₂CH₂—[O—(CH₂)₂]_(n)—NR⁶—,—CH₂CH₂—[O—(CH₂)₂]_(n)—, —[—(CH₂)₂—O—]_(n)—, —[O—(CH₂)₂]_(n)—,—[O—CH(CH₃)C(O)]_(n)—, —[C(O)—CH(CH₃)—O]_(n)—, —[O—CH₂C(O)]_(n)—,—[C(O)—CH₂—O]_(n)—, a divalent residue of a fatty acid, a divalentresidue of an unsaturated or saturated mono- or di-carboxylic acid; eachof which is optionally substituted with 1, 2, 3, or 4 substituentsindependently selected from R²¹;

n is independently selected at each instance from 0, 1, 2, 3, 4, 5, 6,7, 8, 9, or 10;

R²¹ is independently at each occurrence selected from the groupconsisting of hydrogen, alkyl, alkenyl, alkynyl, F, Cl, Br, I, hydroxyl,alkoxy, azide, amino, cyano, —NR⁶R⁷, —NR⁸SO₂R³, —NR⁸S(O)R³, haloalkyl,heteroalkyl, aryl, heteroaryl, and heterocycle;

and the remaining variables are as defined herein.

In one embodiment Linker^(A) is bond and Linker^(B) is

In one embodiment Linker^(B) is bond and Linker^(A)is

In one embodiment, a divalent residue of an amino acid is selected from

wherein the amino acid can be oriented in either direction and whereinthe amino acid can be in the L— or D—form or a mixture thereof.

In one embodiment, a divalent residue of a dicarboxylic acid isgenerated from a nucleophilic addition reaction:

Non-limiting embodiments of a divalent residue of a dicarboxylic acidgenerated from a nucleophilic addition reaction include:

In one embodiment, a divalent residue of a dicarboxylic acid isgenerated from a condensation reaction:

Non-limiting embodiments of a divalent residue of a dicarboxylic acidgenerated from a condensation include:

Non-limiting embodiments of a divalent residue of a saturateddicarboxylic acid include:

Non-limiting embodiments of a divalent residue of a saturateddicarboxylic acid include:

Non-limiting embodiments of a divalent residue of a saturatedmonocarboxylic acid is selected from butyric acid (—OC(O)(CH₂)₂CH₂—),caproic acid (—OC(O)(CH₂)₄CH₂—), caprylic acid (—OC(O)(CH₂)₅CH₂—),capric acid (—OC(O)(CH₂)₈CH₂—), lauric acid (—OC(O)(CH₂)₁₀CH₂—),myristic acid (—OC(O)(CH₂)₁₂CH₂—), pentadecanoic acid(—OC(O)(CH₂)₁₃CH₂—), palmitic acid (—OC(O)(CH₂)₁₄CH₂—), stearic acid(—OC(O)(CH₂)₁₆CH₂—), behenic acid (—OC(O)(CH₂)₂₀CH₂—), and lignocericacid (—OC(O)(CH₂)₂₂CH₂—);

Non-limiting embodiments of a divalent residue of a fatty acid includeresidues selected from linoleic acid, palmitoleic acid, vaccenic acid,paullinic acid, oleic acid, elaidic acid, gondoic acid, gadoleic acid,nervonic acid, myristoleic acid, and erucic acid:

Non-limiting embodiments of a divalent residue of a fatty acid isselected from linoleic acid (—C(O)(CH₂)₇(CH)₂CH₂(CH)₂(CH₂)₄CH₂—),docosahexaenoic acid (—C(O)(CH₂)₂(CHCHCH₂)₆CH₂—), eicosapentaenoic acid(—C(O)(CH₂)₃(CHCHCH₂)₅CH₂—), alpha-linolenic acid(—C(O)(CH₂)₇(CHCHCH₂)₃CH₂—) stearidonic acid(—C(O)(CH₂)₄(CHCHCH₂)₄CH₂—), y-linolenic acid(—C(O)(CH₂)₄(CHCHCH₂)₃(CH₂)₃CH₂—), arachidonic acid(—C(O)(CH₂)₃,(CHCHCH₂)₄(CH₂)₄CH₂—), docosatetraenoic acid(—C(O)(CH₂)₅(CHCHCH₂)₄(CH₂)₄CH₂—), palmitoleic acid(—C(O)(CH₂)₇CHCH(CH₂)₅CH₂—), vaccenic acid (—C(O)(CH₂)₉CHCH(CH₂)₅CH₂—),paullinic acid (—C(O)(CH₂)₁₁CHCH(CH₂)₅CH₂—), oleic acid(—C(O)(CH₂)₇CHCH(CH₂)₇CH₂—), elaidic acid (—C(O)(CH₂)₇CHCH(CH₂)₇CH₂—),gondoic acid (—C(O)(CH₂)₉CHCH(CH₂)₇CH₂—), gadoleic acid(—C(O)(CH₂)₇CHCH(CH₂)₉CH₂—), nervonic acid (—C(O)(CH₂)₁₃CHCH(CH₂)₇CH₂—),mead acid (—C(O)(CH₂)₃(CHCHCH₂)₃(CH₂)₆CH₂—), myristoleic acid(—C(O)(CH₂)₇CHCH(CH₂)₃CH₂—), and erucic acid(—C(O)(CH₂)₁₁CHCH(CH₂)₇CH₂—).

In certain embodiments Linker^(A) is selected from:

wherein:

R²² is independently at each occurrence selected from the groupconsisting of alkyl, —C(O)N—, —NC(O)—, —N—, —C(R²¹)—, —P(O)O—, —P(O)—,—P(O)(NR⁶R⁷)N—, alkenyl, haloalkyl, aryl, heterocycle, and heteroaryl,each of which is optionally substituted with 1, 2, 3, or 4 substituentsindependently selected from R²¹;

and the remaining variables are as defined herein.

In certain embodiments Linker^(D) is selected from:

wherein:

R³² is independently at each occurrence selected from the groupconsisting of alkyl, N⁺ X⁻, —C—, alkenyl, haloalkyl, aryl, heterocycle,and heteroaryl, each of which is optionally substituted with 1, 2, 3, or4 substituents independently selected from R²¹;

X⁻ is an anionic group, for example Br⁻ or Cl⁻ and

all other variables are as defined herein.

In certain embodiments Linker^(A) is selected from:

wherein each heteroaryl, heterocycle, cycloalkyl, and aryl canoptionally be substituted with 1, 2, 3, or 4 of any combination ofhalogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl,as allowed by valence.

In certain embodiments Linker^(A) is selected from:

wherein each heteroaryl, heterocycle, cycloalkyl, and aryl canoptionally be substituted with 1, 2, 3, or 4 of any combination ofhalogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl,as allowed by valence.

In certain embodiments Linker^(A) is selected from:

wherein each heteroaryl, heterocycle, cycloalkyl, and aryl canoptionally be substituted with 1, 2, 3, or 4 of any combination ofhalogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl,as allowed by valence.

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B), Linker^(C), or Linker^(D) is selectedfrom:

wherein tt is independently selected from 1, 2, or 3 and ss is 3 minustt.

In certain embodiments Linker^(B), Linker^(C), or Linker^(D) is selectedfrom:

wherein tt and ss are as defined herein.

In certain embodiments Linker^(B), Linker^(C), or Linker^(D) is selectedfrom:

wherein each heteroaryl, heterocycle, cycloalkyl, and aryl canoptionally be substituted with 1, 2, 3, or 4 of any combination ofhalogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl,as allowed by valence; and tt and ss are as defined herein.

In certain embodiments Linker^(B), Linker^(C), or Linker^(D) is selectedfrom:

wherein each heteroaryl, heterocycle, cycloalkyl, and aryl canoptionally be substituted with 1, 2, 3, or 4 of any combination ofhalogen, alkyl, haloalkyl, aryl, heteroaryl, heterocycle, or cycloalkyl,as allowed by valence; and tt and ss are as defined herein.

In certain embodiments Linker^(B) Linker^(C) or Linker” is selectedfrom:

wherein each heteroaryl and aryl can optionally be substituted with 1,2, 3, or 4 of any combination of halogen, alkyl, haloalkyl, aryl,heteroaryl, heterocycle, or cycloalkyl, as allowed by valence; and ttand ss are as defined herein.

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Lin e is selected from:

In certain embodiments Linker^(D) is selected from

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments, the Linker^(A)is selected from

In certain embodiments, the Linker A is selected from

In certain embodiments, the Linker^(A) is selected from

In certain embodiments, the Linker^(A) is selected from

wherein each is optionally substituted with 1, 2, 3, or 4 substituentssubstituent selected from R²¹.

In certain embodiments Linker A is selected from:

In certain embodiments Linker A is selected from.

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments, the Linker^(B) is selected from

In certain embodiments, the Linker^(A) is selected from

In certain embodiments, the Linker^(A) is selected from

wherein each is optionally substituted with 1, 2, 3, or 4 substituentssubstituent selected from R²¹.

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(A) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B) is selected from:

In certain embodiments Linker^(B)-Linker^(A) is selected from:

In certain embodiments Linker-˜Linker^(A) is selected from:

In certain embodiments, the Linker^(C) is selected from

In certain embodiments, the Linker^(C) is selected from

In certain embodiments, the Linker^(C) is selected from

In certain embodiments, the Linker^(A) is selected from

In certain embodiments, the Linker^(C) is selected from

In certain embodiments, the Linker^(C) is selected from

wherein each is optionally substituted with 1, 2, 3, or 4 substituentssubstituent selected from R²¹.

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C) is selected from:

In certain embodiments Linker^(C)-(Linker^(A))₂ is selected from:

In certain embodiments Linker^(C)-(Linker^(A))₂ is selected from:

In certain embodiments Linker^(C)-(Linker^(A))₂ is selected from:

In certain embodiments Linker^(C) (Linker)₂ is selected from:

In certain embodiments, the Linker^(D) is selected from

In certain embodiments, the Linker^(D) is selected from

In certain embodiments, the Linker^(D) is selected from

wherein each is optionally substituted with 1, 2, 3, or 4 substituentsare selected from R²¹.

In certain embodiments, Linker^(B)-(Linker^(A)) is selected from

In certain embodiments, Linker^(C)-(Linker^(A)) is selected from

In certain embodiments, Linker^(D)-(Linker^(A)) is selected from

V. Compound Terminology

Compounds are described using standard nomenclature. Unless definedotherwise, all technical and scientific terms used herein have the samemeaning as is commonly understood by one of skill in the art to whichthis invention belongs.

The compounds in any of the Formulas described herein include asseparate embodiments enantiomers, diastereomers, tautomers, racemates,rotamers or mixtures thereof, as if each is specifically described,unless otherwise indicated or otherwise excluded by context.

The terms “a” and “an” do not denote a limitation of quantity, butrather denote the presence of at least one of the referenced item. Theterm “or” means “and/or”. Recitation of ranges of values are merelyintended to serve as a shorthand method of referring individually toeach separate value falling within the range, unless otherwise indicatedherein, and each separate value is incorporated into the specificationas if it were individually recited herein. The endpoints of all rangesare included within the range and independently combinable. All methodsdescribed herein can be performed in a suitable order unless otherwiseindicated herein or otherwise clearly contradicted by context. The useof examples, or exemplary language (e.g., “such as”), is intended merelyto better illustrate the invention and does not pose a limitation on thescope of the invention unless otherwise claimed. Unless definedotherwise, technical and scientific terms used herein have the samemeaning as is commonly understood by one of skill in the art to whichthis invention belongs.

The present invention includes compounds with at least one desiredisotopic substitution of an atom, at an amount above the naturalabundance of the isotope, i.e., enriched.

Examples of isotopes that can be incorporated into compounds of theinvention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, fluorine, and chlorine, such as ²H, ³H, ¹¹C, ¹³C, ¹⁴C, ¹⁵N,¹⁷O, ¹⁸O, ¹⁸F ³¹P, ³²P, ³⁵S, ³⁶CI, and ¹²⁵I respectively. In oneembodiment, isotopically labelled compounds can be used in metabolicstudies (with, for example ¹⁴C), reaction kinetic studies (with, forexample ²H or ³H), detection or imaging techniques, such as positronemission tomography (PET) or single-photon emission computed tomography(SPECT) including drug or substrate tissue distribution assays, or inradioactive treatment of patients. For example, a ¹⁸F labeled compoundmay be desirable for PET or SPECT studies. Isotopically labeledcompounds of this invention and prodrugs thereof can generally beprepared by carrying out the procedures disclosed in the schemes or inthe examples and preparations described below by substituting a readilyavailable isotopically labeled reagent for a non-isotopically labeledreagent.

By way of general example and without limitation, isotopes of hydrogen,for example, deuterium (²H) and tritium (3H) may optionally be usedanywhere in described structures that achieves the desired result.Alternatively, or in addition, isotopes of carbon, e.g., ¹³C and ¹⁴C,may be used. In one embodiment, the isotopic substitution is replacinghydrogen with a deuterium at one or more locations on the molecule toimprove the performance of the drug, for example, the pharmacodynamics,pharmacokinetics, biodistribution, half-life, stability, AUC, Tmax,Cmax, etc.

For example, the deuterium can be bound to carbon in a location of bondbreakage during metabolism (an a-deuterium kinetic isotope effect) ornext to or near the site of bond breakage (a β-deuterium kinetic isotopeeffect).

Isotopic substitutions, for example deuterium substitutions, can bepartial or complete. Partial deuterium substitution means that at leastone hydrogen is substituted with deuterium. In certain embodiments, theisotope is 80, 85, 90, 95 or 99% or more enriched in an isotope at anylocation of interest. In certain embodiments deuterium is 80, 85, 90, 95or 99% enriched at a desired location. Unless otherwise stated, theenrichment at any point is above natural abundance, and in an embodimentis enough to alter a detectable property of the drug in a human.

In one embodiment, the substitution of a hydrogen atom for a deuteriumatom occurs within any variable group. For example, when any variablegroup is, or contain for example through substitution, methyl, ethyl, ormethoxy, the alkyl residue may be deuterated (in nonlimitingembodiments, CDH₂, CD₂H, CD₃, CD₂CD₃, CHDCH₂D, CH₂CD₃, CHDCHD₂, OCDH₂,OCD₂H, or OCD₃ etc.). In certain other embodiments, a variable group hasa “'”or an “a” designation, which in one embodiment can be deuterated.In certain other embodiments, when two substituents of the central corering are combined to form a cyclopropyl ring, the unsubstitutedmethylene carbon may be deuterated.

The compound of the present invention may form a solvate with solvents(including water). Therefore, in one embodiment, the invention includesa solvated form of the active compound. The term “solvate” refers to amolecular complex of a compound of the present invention (including asalt thereof) with one or more solvent molecules. Nonlimiting examplesof solvents are water, ethanol, dimethyl sulfoxide, acetone and othercommon organic solvents. The term “hydrate” refers to a molecularcomplex comprising a compound of the invention and water.Pharmaceutically acceptable solvates in accordance with the inventioninclude those wherein the solvent of crystallization may be isotopicallysubstituted, e.g. D₂O, d₆-acetone, d₆-DMSO. A solvate can be in a liquidor solid form.

A dash (“-”) that is not between two letters or symbols is used toindicate a point of attachment for a substituent. For example, —(C═O)NH₂is attached through carbon of the keto (C═O) group.

The term “substituted”, as used herein, means that any one or morehydrogens on the designated atom or group is replaced with a moietyselected from the indicated group, provided that the designated atom'snormal valence is not exceeded and the resulting compound is stable. Forexample, when the substituent is oxo (i.e., ═O) then two hydrogens onthe atom are replaced. For example a pyridyl group substituted by oxo isa pyridone. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds oruseful synthetic intermediates.

“Alkyl” is a branched, straight chain, or cyclic saturated aliphatichydrocarbon group. In one embodiment, the alkyl contains from 1 to about12 carbon atoms, more generally from 1 to about 6 carbon atoms, from 1to about 4 carbon atoms, or from 1 to 3 carbon atoms. In one embodiment,the alkyl contains from 1 to about 8 carbon atoms. In certainembodiments, the alkyl is C₁-C₂, C₁-C₃, C₁-C₄, C₁-C₅ or C₁-C₆. Thespecified ranges as used herein indicate an alkyl group which isconsidered to explicitly disclose as individual species each member ofthe range described as a unique species. For example, the term C₁-C₆alkyl as used herein indicates a straight or branched alkyl group havingfrom 1, 2, 3, 4, 5, or 6 carbon atoms and also a carbocyclic alkyl groupof 3, 4, 5, or 6 carbon atoms and is intended to mean that each of theseis described as an independent species. For example, the term C₁-C₄alkylas used herein indicates a straight or branched alkyl group having from1, 2, 3, or 4 carbon atoms and is intended to mean that each of these isdescribed as an independent species. When C₀-C_(n) alkyl is used hereinin conjunction with another group, for example, (C₃.C₇cycloalkyl)C₀-C₄alkyl, or —C₀-C₄alkyl(C₃-C₇cycloalkyl), the indicated group, in thiscase cycloalkyl, is either directly bound by a single covalent bond(C₀alkyl), or attached by an alkyl chain in this case 1, 2, 3, or 4carbon atoms. Alkyls can also be attached via other groups such asheteroatoms as in —O—C₀-C₄alkyl(C₃-C₇cycloalkyl). Examples of alkylinclude, but are not limited to, methyl, ethyl, n-propyl, isopropyl,n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, isopentyl, tert-pentyl,neopentyl, n-hexyl, 2-methylpentane, 3-methylpentane,2,2-dimethylbutane, 2,3-dimethylbutane, and hexyl.

When a term is used that includes “alk” it should be understood that“cycloalkyl” or “carbocyclic” can be considered part of the definition,unless unambiguously excluded by the context. For example and withoutlimitation, the terms alkyl, alkenyl, alkynyl, alkoxy, alkanoyl,alkenloxy, haloalkyl, etc. can all be considered to include the cyclicforms of alkyl, unless unambiguously excluded by context.

“Alkenyl” is a branched or straight chain aliphatic hydrocarbon grouphaving one or more carbon-carbon double bonds that may occur at a stablepoint along the chain. Nonlimiting examples are C₂-C₈alkenyl,C₂-C₇alkenyl, C₂-C₆alkenyl, C₂-C₅alkenyl and C₂-C₄alkenyl. The specifiedranges as used herein indicate an alkenyl group having each member ofthe range described as an independent species, as described above forthe alkyl moiety. Examples of alkenyl include, but are not limited to,ethenyl and propenyl.

“Alkynyl” is a branched or straight chain aliphatic hydrocarbon grouphaving one or more carbon-carbon triple bonds that may occur at anystable point along the chain, for example, C₂-C₈alkynyl or C₂-C₆alkynyl.The specified ranges as used herein indicate an alkynyl group havingeach member of the range described as an independent species, asdescribed above for the alkyl moiety. Examples of alkynyl include, butare not limited to, ethynyl, propynyl, 1-butynyl, 2-butynyl, 3-butynyl,1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl,3-hexynyl, 4-hexynyl and 5-hexynyl.

“Alkoxy” is an alkyl group as defined above covalently bound through anoxygen bridge (—O—). Examples of alkoxy include, but are not limited to,methoxy, ethoxy, n-propoxy, i-propoxy, n-butoxy, 2-butoxy, t-butoxy,n-pentoxy, 2-pentoxy, 3-pentoxy, isopentoxy, neopentoxy, n-hexoxy,2-hexoxy, 3-hexoxy, and 3-methylpentoxy. Similarly an “alkylthio” or a“thioalkyl” group is an alkyl group as defined above with the indicatednumber of carbon atoms covalently bound through a sulfur bridge (—S—).In one embodiment, the alkoxy group is optionally substituted asdescribed above.

“Haloalkyl” indicates both branched and straight-chain alkyl groupssubstituted with 1 or more halogen atoms, up to the maximum allowablenumber of halogen atoms. Examples of haloalkyl include, but are notlimited to, trifluoromethyl, monofluoromethyl, difluoromethyl,2-fluoroethyl, and penta-fluoroethyl.

“Aryl” indicates an aromatic group containing only carbon in thearomatic ring or rings. In one embodiment, the aryl group contains 1 to3 separate or fused rings and is 6 to 14 or 18 ring atoms, withoutheteroatoms as ring members. The term “aryl” includes groups where asaturated or partially unsaturated carbocycle group is fused with anaromatic ring. The term “aryl” also includes groups where a saturated orpartially unsaturated heterocycle group is fused with an aromatic ringso long as the attachment point is the aromatic ring. Such compounds mayinclude aryl rings fused to a 4 to 7 or a 5 to 7-membered saturated orpartially unsaturated cyclic group that optionally contains 1, 2 or 3heteroatoms independently selected from N, O, B, P, Si and S, to form,for example, a 3,4-methylenedioxyphenyl group. Aryl groups include, forexample, phenyl and naphthyl, including 1-naphthyl and 2-naphthyl. Inone embodiment, aryl groups are pendant. An example of a pendant ring isa phenyl group substituted with a phenyl group.

The term “heterocycle” refers to saturated and partially saturatedheteroatom-containing ring radicals, where the heteroatoms may beselected from N, S, and O. The term “heterocycle” includes monocyclic3-12 membered rings, as well as bicyclic 5-16 membered ring systems(which can include fused, bridged, or spiro, bicyclic ring systems). Itdoes not include rings containing-0—O— or —S—S— portions. Examples ofsaturated heterocycle groups include saturated 4- to 7-memberedmonocyclic groups containing 1 to 4 nitrogen atoms [e.g., pyrrolidinyl,imidazolidinyl, piperidinyl, pyrrolinyl, azetidinyl, piperazinyl, andpyrazolidinyl]; saturated 4 to 6-membered monocyclic groups containing 1to 2 oxygen atoms and 1 to 3 nitrogen atoms [e.g., morpholinyl];saturated 3 to 6-membered heteromonocyclic group containing 1 to 2sulfur atoms and 1 to 3 nitrogen atoms [e.g., thiazolidinyl]. Examplesof partially saturated heterocycle radicals include but are not limitedto, dihydrothienyl, dihydropyranyl, dihydrofuryl, and dihydrothiazolyl.Examples of partially saturated and saturated heterocycle groups includebut are not limited to, pyrrolidinyl, imidazolidinyl, piperidinyl,pyrrolinyl, pyrazolidinyl, piperazinyl, morpholinyl, tetrahydropyranyl,thiazolidinyl, dihydrothienyl, 2,3-dihydro-benzo[1,4]dioxanyl,indolinyl, isoindolinyl, dihydrobenzothienyl, dihydrobenzofuryl,isochromanyl, chromanyl, 1,2-dihydroquinolyl, 1,2,3,4-tetrahydro-isoquinolyl, 1,2,3,4-tetrahydro-quinolyl,2,3,4,4a,9,9a-hexahydro-1H-3-aza-fluorenyl,5,6,7-trihydro-1,2,4-triazolo[3,4-a]isoquinolyl,3,4-dihydro-2H-benzo[1,4]oxazinyl, benzo[1,4]dioxanyl, 2,3-dihydro-1H-1λ-benzo[d]isothiazol-6-yl, dihydropyranyl, dihydrofuryl anddihydrothiazolyl. “Bicyclic heterocycle” includes groups wherein theheterocyclic radical is fused with an aryl radical wherein the point ofattachment is the heterocycle ring. “Bicyclic heterocycle” also includesheterocyclic radicals that are fused or bridged with a carbocycleradical. For example partially unsaturated condensed heterocyclic groupcontaining 1 to 5 nitrogen atoms, for example, indoline, isoindoline,partially unsaturated condensed heterocyclic group containing 1 to 2oxygen atoms and 1 to 3 nitrogen atoms, partially unsaturated condensedheterocyclic group containing 1 to 2 sulfur atoms and 1 to 3 nitrogenatoms, and saturated condensed heterocyclic group containing 1 to 2oxygen or sulfur atoms.

Non-limiting examples of bicyclic heterocycles include:

Unless otherwise drawn or clear from the context, the term “bicyclicheterocycle” includes cis and trans diastereomers. Non-limiting examplesof chiral bicyclic heterocycles include:

In certain alternative embodiments the term “heterocycle” refers tosaturated and partially saturated heteroatom-containing ring radicals,where the heteroatoms may be selected from N, S, O, B, Si, and P.

“Heteroaryl” refers to a stable monocyclic, bicyclic, or multicyclicaromatic ring which contains from 1 to 3, or in some embodiments from 1,2, or 3 heteroatoms selected from N, O, S, B, and P (and typicallyselected from N, O, and S) with remaining ring atoms being carbon, or astable bicyclic or tricyclic system containing at least one 5, 6, or 7membered aromatic ring which contains from 1 to 3, or in someembodiments from 1 to 2, heteroatoms selected from N, O, S, B or P withremaining ring atoms being carbon. In one embodiment, the onlyheteroatom is nitrogen. In one embodiment, the only heteroatom isoxygen. In one embodiment, the only heteroatom is sulfur. Monocyclicheteroaryl groups typically have from 5 or 6 ring atoms. In someembodiments bicyclic heteroaryl groups are 8- to 10-membered heteroarylgroups, that is, groups containing 8 or 10 ring atoms in which one 5, 6,or 7-member aromatic ring is fused to a second aromatic or non-aromaticring wherein the point of attachment is the aromatic ring. When thetotal number of S and O atoms in the heteroaryl group exceeds 1, theseheteroatoms are not adjacent to one another. In one embodiment, thetotal number of S and O atoms in the heteroaryl group is not more than2. In another embodiment, the total number of S and O atoms in thearomatic heterocycle is not more than 1. Examples of heteroaryl groupsinclude, but are not limited to, pyridinyl (including, for example,2-hydroxypyridinyl), imidazolyl, imidazopyridinyl, pyrimidinyl(including, for example, 4-hydroxypyrimidinyl), pyrazolyl, triazolyl,pyrazinyl, furyl, thienyl, isoxazolyl, thiazolyl, oxadiazolyl, oxazolyl,isothiazolyl, pyrrolyl, quinolinyl, isoquinolinyl,tetrahydroisoquinolinyl, indolyl, benzimidazolyl, benzofuranyl,cinnolinyl, indazolyl, indolizinyl, phthalazinyl, pyridazinyl,triazinyl, isoindolyl, pteridinyl, purinyl, oxadiazolyl, triazolyl,thiadiazolyl, thiadiazolyl, furazanyl, benzofurazanyl, benzothiophenyl,benzothiazolyl, benzoxazolyl, quinazolinyl, quinoxalinyl,naphthyridinyl, tetrahydrofuranyl, and furopyridinyl. Heteroaryl groupsare optionally substituted independently with one or more substituentsdescribed herein.

“Heteroaryloxy” is a heteroaryl group as described bound to the group itsubstituted via an oxygen, —O—, linker.

“Heteroarylalkyl” is an alkyl group as described herein substituted witha heteroaryl group as described herein.

“Arylalkyl” is an alkyl group as described herein substituted with anaryl group as described herein.

“Heterocycloalkyl” is an alkyl group as described herein substitutedwith a heterocyclo group as described herein.

The term “heteroalkyl” refers to an alkyl, alkenyl, alkynyl, orhaloalkyl moiety as defined herein wherein a CH₂ group is eitherreplaced by a heteroatom or a carbon atom is substituted with aheteroatom for example, an amine, carbonyl, carboxy, oxo, thio,phosphate, phosphonate, nitrogen, phosphorus, silicon, or boron. In oneembodiment, the only heteroatom is nitrogen. In one embodiment, the onlyheteroatom is oxygen. In one embodiment, the only heteroatom is sulfur.In one embodiment, “heteroalkyl” is used to indicate a heteroaliphaticgroup (cyclic, acyclic, substituted, unsubstituted, branched orunbranched) having 1-20 carbon atoms. Nonlimiting examples ofheteroalkyl moieties include polyethylene glycol, polyalkylene glycol,amide, polyamide, polylactide, polyglycolide, thioether, ether,alkyl-heterocycle-alkyl, —O—alkyl—O—alkyl, alkyl—O—haloalkyl, etc.

When a compound moiety is “optionally substituted” it may be substitutedas allowed by valence with one or more groups selected from alkyl(including C₁-C₄alkyl), alkenyl (including C₂-C₄alkenyl), alkynyl(including C₂-C₄alkynyl), haloalkyl (including C₁-C₄haloalkyl), —OR⁶, F,Cl, Br, I, —NR⁶R⁷, heteroalkyl, cyano, nitro, C(O)R³

wherein the optional substituent is selected such that a stable compoundresults. For example

could be substituted with 1 or 2 groups independently selected fromalkyl, alkenyl, alkynyl, haloalkyl, —OR⁶, F, Cl, Br, I, —NR⁶R⁷,heteroalkyl, cyano, nitro, C(O)R³ so long as a stable compound resultsbut only one group selected from

so long as a stable compound results.

on the other hand could only be substituted with 1 or 2 groups selectedfrom

Non-limiting examples of optionally substituted CH₂ groups include:

Non-limiting examples of optionally substituted —S— groups include:

A “dosage form” means a unit of administration of an active agent.Examples of dosage forms include tablets, capsules, injections,suspensions, liquids, emulsions, implants, particles, spheres, creams,ointments, suppositories, inhalable forms, transdermal forms, buccal,sublingual, topical, gel, mucosal, subcutaneous, intramuscular,parenteral, systemic, intravenous, and the like. A “dosage form” canalso include an implant for controlled delivery.

“Pharmaceutical compositions” are compositions comprising at least oneactive agent, and at least one other substance, such as a carrier. Thepresent invention includes pharmaceutical compositions of the describedcompounds.

“Pharmaceutical combinations” are combinations of at least two activeagents which may be combined in a single dosage form or providedtogether in separate dosage forms with instructions that the activeagents are to be used together to treat any disorder described herein.

A “pharmaceutically acceptable salt” is a derivative of the disclosedcompound in which the parent compound is modified by making an inorganicor organic, pharmaceutically acceptable, acid or base addition saltsthereof. The salts of the present compounds can be synthesized from aparent compound that contains a basic or acidic moiety by conventionalchemical methods. Generally, such salts can be prepared by reacting freeacid forms of these compounds with a stoichiometric amount of theappropriate base (such as Na, Ca, Mg, or K hydroxide, carbonate,bicarbonate, or the like), or by reacting free base forms of thesecompounds with a stoichiometric amount of the appropriate acid. Suchreactions are typically carried out in water or in an organic solvent,or in a mixture of the two. Salts of the present compounds furtherinclude solvates of the compounds and of the compound salts.

Examples of pharmaceutically acceptable salts include, but are notlimited to, mineral or organic acid salts of basic residues such asamines; alkali or organic salts of acidic residues such as carboxylicacids; and the like. The pharmaceutically acceptable salts include saltswhich are acceptable for human consumption and the quaternary ammoniumsalts of the parent compound formed, for example, from inorganic ororganic acids. Examples, of such salts include those derived frominorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic,phosphoric, nitric and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic,sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic,ethane disulfonic, oxalic, isethionic, HOOC—(CH₂)₁₋₄-COOH, and the like,or using a different acid that produces the same counterion. Lists ofadditional suitable salts may be found, e.g., in Remington'sPharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa.,p. 1418 (1985).

The term “carrier” applied to pharmaceutical compositions/combinationsof the invention refers to a diluent, excipient, or vehicle with whichan active compound is provided.

A “pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition/combination that isgenerally safe, acceptable for human consumption, and neitherbiologically nor otherwise inappropriate for administration to a host,typically a human. In one embodiment, an excipient is used that isacceptable for veterinary use.

A “patient” or “host” or “subject” is a human or non-human animal inneed of treatment or prevention of any of the disorders as specificallydescribed herein. Typically the host is a human.

A “patient” or “host” or “subject” also refers to for example, a mammal,primate (e.g., human), cow, sheep, goat, horse, dog, cat, rabbit, rat,mice, bird and the like.

A “therapeutically effective amount” of a compound, pharmaceuticalcomposition, or combination of this invention means an amount effective,when administered to a host, that provides a therapeutic benefit such asan amelioration of symptoms or reduction or dimunition of the diseaseitself. In another aspect, a preventative amount can be administeredthat prevents or minimizes the risk of the disease mediated by theExtracellular Target Protein.

Embodiments of “alkyl”

In one embodiment “alkyl” is a C₁-C₁₀alkyl, C₁-C₉alkyl, C₁-C₈alkyl,C₁-C₇alkyl, C₁-C₆alkyl, C₁-C₈alkyl, C₁-C₄alkyl, C₁-C₃alkyl, orC₁-C₂alkyl.

In one embodiment “alkyl” has one carbon.

In one embodiment “alkyl” has two carbons.

In one embodiment “alkyl” has three carbons.

In one embodiment “alkyl” has four carbons.

In one embodiment “alkyl” has five carbons.

In one embodiment “alkyl” has six carbons.

Non-limiting examples of “alkyl” include: methyl, ethyl, propyl, butyl,pentyl, and hexyl.

Additional non-limiting examples of “alkyl” include: isopropyl,isobutyl, isopentyl, and isohexyl.

Additional non-limiting examples of “alkyl” include: sec-butyl,sec-pentyl, and sec-hexyl.

Additional non-limiting examples of “alkyl” include: tert-butyl,tert-pentyl, and tert-hexyl.

Additional non-limiting examples of “alkyl” include: neopentyl,3-pentyl, and active pentyl.

In an alternative embodiment the “alkyl” group is optionallysubstituted.

In an alternative embodiment the “alkenyl” group is optionallysubstituted.

In an alternative embodiment the “alkynyl” group is optionallysubstituted.

Embodiments of “Haloalkyl”

In one embodiment “haloalkyl” is a C₁-C₁₀haloalkyl, C₁-C₉haloalkyl,C₁-C⁸haloalkyl, C₁-C₇haloalkyl, C₁-C⁶haloalkyl, C₁-C⁵haloalkyl,C₁-C⁴haloalkyl, C₁-C³haloalkyl, and C₁-C₂haloalkyl.

In one embodiment “haloalkyl” has one carbon.

In one embodiment “haloalkyl” has one carbon and one halogen.

In one embodiment “haloalkyl” has one carbon and two halogens.

In one embodiment “haloalkyl” has one carbon and three halogens.

In one embodiment “haloalkyl” has two carbons.

In one embodiment “haloalkyl” has three carbons.

In one embodiment “haloalkyl” has four carbons.

In one embodiment “haloalkyl” has five carbons.

In one embodiment “haloalkyl” has six carbons.

Non-limiting examples of “haloalkyl” include:

Additional non-limiting examples of “haloalkyl” include:

Additional non-limiting examples of “haloalkyl” include

Additional non-limiting examples of “haloalkyl” include:

Embodiments of “Heteroaryl”

Non-limiting examples of 5 membered “heteroaryl” groups include pyrrole,furan, 5 thiophene, pyrazole, imidazole, triazole, isoxazole, oxazole,oxadiazole, oxatriazole, isothiazole, thiazole, thiadiazole, andthiatriazole.

Additional non-limiting examples of 5 membered “heteroaryl” groupsinclude:

In one embodiment “heteroaryl” is a 6 membered aromatic group containing1, 2, or 3 nitrogen atoms (i.e. pyridinyl, pyridazinyl, triazinyl,pyrimidinyl, and pyrazinyl).

Non-limiting examples of 6 membered “heteroaryl” groups with 1 or 2nitrogen atoms include:

In one embodiment “heteroaryl” is a 9 membered bicyclic aromatic groupcontaining 1 or 2 atoms selected from nitrogen, oxygen, and sulfur.

Non-limiting examples of “heteroaryl” groups that are bicyclic includeindole, benzofuran, isoindole, indazole, benzimidazole, azaindole,azaindazole, purine, isobenzofuran, benzothiophene, benzoisoxazole,benzoisothiazole, benzooxazole, and benzothiazole.

Additional non-limiting examples of “heteroaryl” groups that arebicyclic include:

Additional non-limiting examples of “heteroaryl” groups that arebicyclic include:

Additional non-limiting examples of “heteroaryl” groups that arebicyclic include:

In one embodiment “heteroaryl” is a 10 membered bicyclic aromatic groupcontaining 1 or 2 atoms selected from nitrogen, oxygen, and sulfur.

Non-limiting examples of “heteroaryl” groups that are bicyclic includequinoline, isoquinoline, quinoxaline, phthalazine, quinazoline,cinnoline, and naphthyridine.

Additional non-limiting examples of “heteroaryl” groups that arebicyclic include:

Embodiments of “Heterocycle”

In one embodiment “heterocycle” refers to a cyclic ring with onenitrogen and 3, 4, 5, 6, 7, or 8 carbon atoms.

In one embodiment “heterocycle” refers to a cyclic ring with onenitrogen and one oxygen and 3, 4, 5, 6, 7, or 8 carbon atoms.

In one embodiment “heterocycle” refers to a cyclic ring with twonitrogens and 3, 4, 5, 6, 7, or 8 carbon atoms.

In one embodiment “heterocycle” refers to a cyclic ring with one oxygenand 3, 4, 5, 6, 7, or 8 carbon atoms.

In one embodiment “heterocycle” refers to a cyclic ring with one sulfurand 3, 4, 5, 6, 7, or 8 carbon atoms.

Non-limiting examples of “heterocycle” include aziridine, oxirane,thiirane, azetidine, 1,3-diazetidine, oxetane, and thietane.

Additional non-limiting examples of “heterocycle” include pyrrolidine,3-pyrroline, 2-pyrroline, pyrazolidine, and imidazolidine.

Additional non-limiting examples of “heterocycle” includetetrahydrofuran, 1,3-dioxolane, tetrahydrothiophene, 1,2-oxathiolane,and 1,3-oxathiolane.

Additional non-limiting examples of “heterocycle” include piperidine,piperazine, tetrahydropyran, 1,4-dioxane, thiane, 1,3-dithiane,1,4-dithiane, morpholine, and thiomorpholine.

Additional non-limiting examples of “heterocycle” include indoline,tetrahydroquinoline, tetrahydroisoquinoline, and dihydrobenzofuranwherein the point of attachment for each group is on the heterocyclicring.

For example

is a “heterocycle” group.

However,

is an “aryl” group.

Non-limiting examples of “heterocycle” also include:

Additional non-limiting examples of “heterocycle” include:

Additional non-limiting examples of “heterocycle” include:

Non-limiting examples of “heterocycle” also include:

Non-limiting examples of “heterocycle” also include:

Additional non-limiting examples of “heterocycle” include:

Additional non-limiting examples of “heterocycle” include:

Aryl

In one embodiment “aryl” is a 6 carbon aromatic group (phenyl).

In one embodiment “aryl” is a 10 carbon aromatic group (naphthyl).

In one embodiment “aryl” is a 6 carbon aromatic group fused to aheterocycle wherein the point of attachment is the aryl ring.Non-limiting examples of “aryl” include indoline, tetrahydroquinoline,tetrahydroisoquinoline, and dihydrobenzofuran wherein the point ofattachment for each group is on the aromatic ring.

For example

is an “aryl” group.

However,

is a “heterocycle” group.

Embodiments of “arylalkyl”

Non-limiting examples of “arylalkyl” include:

In one embodiment “arylalkyl” is

In one embodiment the “arylalkyl” refers to a 2 carbon alkyl groupsubstituted with an aryl group.

Non-limiting examples of “arylalkyl” include:

VI. Extracellular Proteins and Targeting Ligands

A wide range of well-known and characterized extracellular proteins cancause, modulate, or amplify diseases in vivo, such as abnormal cellularproliferation such as tumors and cancer, autoimmune disorders,inflammation and aging-related diseases. For example, extracellularproteins such as growth factors, cytokines, and chemokines bind to cellsurface receptors, often initiate aberrant signaling in multiplediseases such as cancer and inflammation.

The extracellular protein degrader described herein or itspharmaceutically acceptable salt and/or its pharmaceutically acceptablecompositions can be used to treat a disorder which is mediated by theselected Target Protein that binds to the Targeting Ligand. Thedescribed degraders are capable of targeting specific extracellularTarget Proteins that mediate pathological disorders for lysosomaldegradation. The selected extracellular Target Protein may modulate adisorder in a human via a mechanism of action such as modification of abiological pathway, pathogenic signaling, or modulation of a signalcascade or cellular entry. In one embodiment, the Target Protein is aprotein that is not druggable in the classic sense in that it does nothave a binding pocket or an active site that can be inhibited orotherwise bound, and cannot be easily allosterically controlled. Inanother embodiment, the Target Protein is a protein that is druggable inthe classic sense, yet for therapeutic purposes, degradation of theprotein is preferred to inhibition. The extracellular Target Protein isrecruited with a Targeting Ligand, which is a ligand for theextracellular Target Protein. Typically, the Targeting Ligand binds theTarget Protein in a non-covalent fashion. In an alternative embodiment,the Target Protein is covalently bound to the Targeting Ligand in amanner that can be irreversible or reversible.

Accordingly, in some embodiments, a method to treat a host with adisorder mediated by an extracellular Target Protein is provided thatincludes administering an effective amount of a degrader targeting anextracellular protein or its pharmaceutically acceptable salt describedherein to the host, typically a human, optionally in a pharmaceuticallyacceptable composition.

The extracellular Target Protein can be any amino acid sequence to whichthe degrader comprising a Targeting Ligand can be bound which throughdegradation thereof, results in a beneficial therapeutic effect. In oneembodiment, the Target Protein is a non-endogenous peptide such as thatfrom a pathogen or toxin. In another embodiment, the Target Protein canbe an endogenous protein that mediates a disorder. The endogenousprotein can be either the normal form of the protein or an aberrantform. For example, the Target Protein can be an extracellular mutantprotein, or a protein, for example, where a partial, or full,gain-of-function or loss-of-function is encoded by nucleotidepolymorphisms. In some embodiments, the degrader targets the aberrantform of the protein and not the normal form of the protein.

The Targeting Ligand is a ligand which covalently or non-covalentlybinds to a Target Protein which has been selected for lysosomaldegradation. A Targeting Ligand is a small molecule or moiety (forexample a peptide, nucleotide, antibody fragment, aptamer, biomolecule,or other chemical structure) that binds to a Target Protein, and whereinthe Target Protein is a mediator of disease in a host as described indetail below. Exemplary Target Ligands are provided in FIG. 1 .

Anchor Bond The Extracellular Protein Target Ligand (“EPTL”) iscovalently bound to Linker in the ASGPR-binding extracellular proteindegrader compound through the Anchor Bond (which is the chemical bondbetween the EPTL and either Linker B, Linker C or Linker D). This bondcan be placed at any location on the ligand that does not unacceptablydisrupt the ability of the EPTL to bind to the Extracellular ProteinTarget. The Anchor Bond is depicted on the nonlimiting examples ofExtracellular Protein Target Ligands in FIG. 1 as:

A number of exemplary extracellular proteins targeted for medicaltherapy described below have characterizing structural information inthe well-known Protein Data Bank (“PDB”), which is a database for thethree-dimensional structural information for large biological moleculessuch as proteins and nucleic acids. PDB includes x-ray crystallographyand other information submitted by scientists around the world, and isfreely accessible. See for example www.rcsb.org; www.wwpdb.org andwww.uniprot.org. Using the PDB codes for example provided in Section **or in the Data Bank itself, and technical references provided herein orotherwise publicly available, the skilled artisan can determineappropriate locations where the EPTL can be linked through an AnchorBond to Linker B, Linker C or Linker D to the ASGPR-binding moiety. Formany of these proteins, published references describe how a range ofligands bind to the target proteins, and from this information, one candetermine reasonable Anchor Bond locations.

For example, the skilled artisan can use available visualization tools,including those available on the PDB website, to determine where theExtracellular Protein Targeting Ligand docks into to the ExtracellularProtein. The skilled artisan can also import the crystal structure andthe selected Extracellular Protein Targeting Ligand of interest intomodeling software (including for example PyMOL, Glide, Maestro, RasMol,Visual Molecular Dynamics, Jmol, and AutoDock) to determine what portionof the Extracellular Protein Targeting Ligand is bound to theExtracellular Protein. The ASGPR ligand is then bound through the Linkerand the Anchor Bond at a point that does not unduly adversely affectbinding to the extracellular protein.

Non-Limiting Examples of Extracellular Target Proteins

Immunoglobulin A (IgA)

In some embodiments, the Target Protein is human immunoglobulin A(IgA).IgA is an antibody that plays a crucial role in the immune function ofmucous membranes. The amount of IgA produced in association with mucosalmembranes is greater than all other types of antibody combined. IgA hastwo subclasses (IgA1 and IgA2) and can be produced as a monomeric aswell as a dimeric form. The IgA dimeric form is the most prevalent. Inthe blood, IgA interacts with an Fc receptor called FcαRI (or CD89),which is expressed on immune effector cells, to initiate inflammatoryreactions. Ligation of FcαRI by IgA containing immune complexes causesantibody-dependent cell-mediated cytotoxicity (ADCC), degranulation ofeosinophils and basophils, phagocytosis by monocytes, macrophages, andneutrophils, and triggering of respiratory burst activity bypolymorphonuclear leukocytes. Aberrant IgA expression has beenimplicated in a number of autoimmune and immune-mediated disorders,including IgA nephropathy, celiac disease, Henoch-Sconiein purpura(HSP), liner IgA bullous dermatosis, and IgA pemphigus.

The Protein Data Bank website provides the crystal structure of IgA, aswell as the crystal structure of IgA bound to various compoundssearchable by 5E8E (Baglin, T. P., et al., J. Thromb. Haemost., 2016,14: 137-142), and 2QTJ (Bonner, A., et al., J. Immunol., 2008, 180:1008-1018). Additionally, Hatanaka T. et al., provides great insightinto the specificity and high binding affinity of IgA to OPT-1 peptides(J Biol Chem., 2012, 287(51), 43126-43136.).

Representative IgA Targeting Ligands are provided in FIG. 1 .

Additional representative IgA Targeting Ligands include:

SEQ ID NO: 1MLKKIE (Jerlstrom et al. Infect. Immun. 1996 Jul; 64(7):2787-2793;;Opt-1- SEQ ID NO: 2HMVCLAYRGRPVCFAL (Hatanaka et al. J. Biol. Chem. Vol. 287, No. 51, pp. 43126-43136, Dec. 14, 2012); Opt-2- SEQ ID NO: 3HMVCLSYRGRPVCFSL (Hatanaka et al. J. Biol. Chem. Vol. 287, No. 51,pp. 43126-43136, Dec. 14, 2012); Opt-3- SEQ ID NO: 4HQVCLSYRGRPVCFST (Hatanaka et al. J. Biol. Chem. Vol. 287, No. 51,pp. 43126-43136, Dec. 14, 2012); SQE ID NO: 5QMRCLSYKGRRVCLWL (U.S. Pat. No. 9593147); SEQ ID NO: 6KRLCLQYKGSKVCFRL (U.S. Pat. No. 9593147); SEQ ID NO: 7RMRCLTYRGRRVCLEL (U.S. Pat. No. 9593147); SEQ ID NO: 8SMRCLQYRGSRVCLTL (U.S. Pat. No. 9593147); SEQ ID NO: 9HLRCLRYKGTRVCFSL (U.S. Pat. No. 9593147); SEQ ID NO: 10HVRCLSYKGREVCVQL (U.S. Pat. No. 9593147); SEQ ID NO: 11PRMCLFIYKGRRVCIPY (U.S. Pat. No. 9593147); SEQ ID NO: 12HMRCLHYKGRRVCFLL (U.S. Pat. No. 9593147); SEQ ID NO: 13HKRCLHYRGRMVCFLI (U.S. Pat. No. 9593147); SEQ ID NO: 14QKRCLKYKGSRVCFFL (U.S. Pat. No. 9593147); SEQ ID NO: 15HVRCLRYRGKNVCFLL (U.S. Pat. No. 9593147); SEQ ID NO: 16SDVCLRYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 17RDVCLRYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 18HDVCLRYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 19SMVCLRYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 20SAVCLRYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 21SDVCLNYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 22SDVCLHYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 23SDVCLAYRGRPVCFQV (U.S. Pat. No. 9593147); SEQ ID NO: 24SDVCLRYRGRPVCFAV (U.S. Pat. No. 9593147); SEQ ID NO: 25SDVCLRYRGRPVCFQL (U.S. Pat. No. 9593147); SEQ ID NO: 26SDVCLRYRGRPVCFQA (U.S. Pat. No. 9593147); SEQ ID NO: 27HMVCLSYRGRPVCF (US Pub. No. 20150044701); SEQ ID NO: 28HMVCLSYRGRPVCFS (US Pub. No. 20150044701); SEQ ID NO: 29HQVCLSYRGQPVCFSL (US Pub. No. 20150044701); SEQ ID NO: 30HQVCLSYRGRPTCFSL (US Pub. No. 20150044701); SEQ ID NO: 31HQVCLSYRGRPVCYSL (US Pub. No. 20150044701); SEQ ID NO: 32HQVCLSYRGQPVCFST (US Pub. No. 20150044701); SEQ ID NO: 33HQVCLSYRGRPTCFST (US Pub. No. 20150044701); SEQ ID NO: 34HQVCLSYRGQPTCFST (US Pub. No. 20150044701);

Immunoglobulin G (IgG)

In some embodiments, the Target Protein is a human immunoglobulin G(IgG). IgG represents approximately 75% of serum antibodies in humans.IgG is the most common type of antibody found in blood circulation. IgGantibodies are large globular proteins with a molecular weight of about150 kDa made of four peptide chains.[6] It contains two identical γ(gamma) heavy chains of about 50 kDa and two identical light chains ofabout 25 kDa, thus a tetrameric quaternary structure. The two heavychains are linked to each other and to a light chain each by disulfidebonds. The resulting tetramer has two identical halves, which togetherform the Y-like shape. Each end of the fork contains an identicalantigen binding site. The various regions and domains of a typical IgGare depicted in the figure to the left. The Fc regions of IgGs bear ahighly conserved N-glycosylation site at asparagine 297 in the constantregion of the heavy chain. The N-glycans attached to this site arepredominantly core-fucosylated biantennary structures of the complextype. In addition, small amounts of these N-glycans also bear bisectingGlcNAc and α-2,6-linked sialic acid residues. The N-glycan compositionin IgG has been linked to several autoimmune, infectious and metabolicdiseases. In addition, overexpression of IgG4 has been associated withIG4- related diseases, which generally include multiple organs, anddisorders include type 1 autoimmune pancreatitis, interstitialnephritis, Riedel's thyroiditis, Mikulicz's disease, Kuttner's tumor,inflammatory pseudotumors (in various sites of the body), mediastinalfibrosis and some cases of retroperitoneal fibrosis, aortitis,retroperitoneal fibrosis, proximal biliary strictures,tubulointerstitial nephritis, pachymeningitis, pancreatic enlargementand pericarditis.

The Protein Data Bank website provides the crystal structure of IgGsearchable by 1H3X (Krapp, S., et al., J. Mol. Biol., 2003, 325: 979);and 5V43 (Lee, C. H., et al., Nat. Immunol., 2017, 18: 889-898); as wellas the crystal structure of IgG bound to various compounds searchable by5YC5 (Kiyoshi M., et al., Sci. Rep., 2018, 8: 3955-3955); 5XJE (SakaeY., et al., Sci. Rep.,2017, 7: 13780-13780); 5GSQ (Chen, C. L., et al.,ACS Chem. Biol., 2017, 12: 1335-1345); and 1HZH (Saphire E. O., et al.,Science, 2001, 293: 1155-1159). Additionally, Kiyoshi, M., et al.,provides insight into the structural basis for binding of human IgGI toits high-affinity human receptor FcγRI. (Kiyosi M., et al., Nat Commun.,2015, 6, 6866).

Representative IgG Targeting Ligands are provided in FIG. 1 .

Additional representative IgG Targeting Ligands include:

wherein X_(R) is O, S, NH, or N—C₁-C₃ alkyl; and

X_(M) is O, S, NH, or N—C₁-C₃ alkyl.

In other embodiments the IgG Targeting Ligand is selected from:

In some embodiments, the IgG Targeting Ligand is a group according tothe chemical 10 structure:

wherein R^(N02) is a dinitrophenyl group optionally linked through CH₂,S(O), S(O)₂, —S(O)₂O,—OS(O)₂, or OS(O)₂O.

In certain embodiments the IgG Targeting Ligand is selected from:

wherein X¹⁰⁰ is selected from O, CH₂, NH, N—C₁-C₃ alkyl, NC(O)C₁-C₃alkyl, S(O), S(O)₂, —S(O)₂O, —OS(O)₂, or OS(O)₂O.

In some embodiments, the IgG Targeting Ligand is a 3-indoleacetic acidgroup according to the chemical structure:

where k″″ is 1-4 (preferably 2-3, most often 3) or a

group.

In some embodiments, the IgG Targeting Ligand is a peptide. Nonlimitingexamples of IgG Targeting Ligand peptides include: PAM (RTY)₄K₂KG(Fassina, et al, J. Mol. Recognit. 1996, 9, 564-569) SEQ ID NO:35;

D-PAM, wherein the amino acids of the PAM sequence are all D-amino acids(Verdoliva, et al, J. Immunol. Methods, 2002, 271, 77-88) (RTY)₄K₂KG SEQID NO:36; D-PAM-Φ, wherein the amino acids of the PAM sequence are allD-amino acids with further modifications wherein the four N-terminalarginines are acetylated with phenylactic acid (Dinon, et al J. Mol.Recognit. 2011, 24, 1087-1094) (RTY)₄K₂KG SEQ ID NO:37;

SEQ ID NO: 38TWKTSRISIF (Krook, et al,.J. Immunol. Methods 1998, 221, 151-157);SEQ ID NO: 39FGRLVSSIRY (Krook, et al, J. Immunol . Methods 1998, 221, 151-157);SEQ ID NO: 40Fc-III (DCAWHLGELVWCT-NH2) (DeLano et al, Science 2000, 287, 1279-1283)

SEQ ID NO: 41 FCBP-SerDSAWHLGELWST (see WO2014010813); SEQ ID NO: 42DCHKRSFWADNCT (see WO2014010813); SEQ ID NO: 43DCRTQFRPNQTCT (see WO2014010813); SEQ ID NO: 44DCQLCDFWRTRCT (see WO2014010813); SEQ ID NO: 45DCFEDFNEQRTCT (see WO2014010813); SEQ ID NO: 46DCLAKFLKGKDCT (see WO2014010813); SEQ ID NO: 47DCWHRRTHKTFCT (see WO2014010813); SEQ ID NO: 48DCRTIQTRSCT (see WO2014010813); SEQ ID NO: 49DCIKLAQLHSVCT (see WO2014010813); SEQ ID NO: 50DCWRHRNATEWCT (see WO2014010813); SEQ ID NO: 51DCQNWIKDVHKCT (see WO2014010813); SEQ ID NO: 52DCAWHLGELVWCT (see WO2014010813); SEQ ID NO: 53DCAFHLGELVWCT (see WO2014010813); SEQ ID NO: 54DCAYHLGELVWCT (see WO2014010813);

FcBP-1 SEQ ID NO: 55PAWHLGELVWP (Kang, et al, J. Chromatogr. A 2016, 1466, 105-1 12) 

FcBP-2 SEQ ID NO: 56PDCAWHLGELVWCTP (Dias, et al, J . Am. Chem. Soc. 2006, 128, 2726-2732);

Fc-lll-4c SEQ ID NO: 57CDCAWHLGELVWCTC (Gong, et al, Bioconjug. Chem. 2016, 27, 1569-1573) 

SEQ ID NO: 58EPIHRSTLTALL (Ehrlich, et al, J. Biochem. Biophys. Method 2001, 49, 443-454);SEQ ID NO: 59APAR (Camperi, et al, Biotechnol. Lett. 2003, 25, 1545-1548);SEQ ID NO: 60FcRM (CFHH)₂KG (Fc Receptor Mimetic, Verdoliva, et al., ChemBioChem 2005, 6,1242-1253) 

SEQ ID NO: 61 HWRGWV (Yang, et al., J Peptide Res. 2006, 66, 110-137);SEQ ID NO: 62HYFKFD (Yang, et al, J. Chromatogr. A 2009, 1216, 910-918);SEQ ID NO: 63HFRRHL (Menegatti, et al, J. Chromatogr. A 2016, 1445, 93-104);SEQ ID NO: 64HWCitGWV (Menegatti, et al, J. Chromatogr. A 2016, 1445, 93-104);SEQ ID NO: 65 HWmetCitGWmetV (US10,266,566); SEQ ID NO: 66DAAG (Small Synthetic peptide ligand, Lund, et al, J. Chromatogr. A 2012, 1225,158-167); SEQ ID NO: 67DAAG (Small Synthetic peptide ligand, Lund, et al, J. Chromatogr. A 2012, 1225, 158-167); SEQ ID NO: 68cyclo[(Na-Ac) S(A)-RWHYFK-Lact-E] (Menegatti, et al, Anal. Chern. 2013, 85,9229-9237); SEQ ID NO: 69cyclo[(Na-Ac)-Dap(A)-RWHYFK-Lact-E] (Menegatti, et al, Anal. Chern. 2013, 85, 9229-9237); SEQ ID NO: 70cyclo[Link M-WFRHYK] (Menegatti, et al, Biotechnol. Bioeng. 2013, 110, 857-870);SEQ ID NO: 71NKFRGKYK (Sugita, et al, Biochem. Eng. J. 2013, 79, 33-40);SEQ ID NO: 72NARKFYKG (Sugita, et al, Biochem. Eng. J. 2013, 79, 33-40);SEQ ID NO: 73 FYWHCLDE (Zhao, et al, Biochem. Eng. J. 2014, 88, 1-11);SEQ ID NO: 74FYCHWALE (Zhao, et al, J Chromatogr. A 2014, 1355, 107-114);SEQ ID NO: 75FYCHTIDE (Zhao, et al., Z Chromatogr. A 2014, 1359, 100-111);SEQ ID NO: 76Dual 1/3 (FYWHCLDE-FYCHTIDE) (Zhao, et al, J. Chromatogr. A 2014, 1369, 64-72);SEQ ID NO: 77 RRGW (Tsai, et al, Anal. Chem. 2014, 86, 293 1-2938);SEQ ID NO: 78 KHRFNKD (Yoo and Choi, BioChip J. 2015, 10, 88-94);SEQ. ID NO: 79 CPSTHWK (Sun et al. Polymers 2018, 10, 778);SEQ. ID NO: 80 NVQYFAV (Sun et al. Polymers 2018, 10, 778);SEQ. ID NO: 81 ASHTQKS (Sun et al. Polymers 2018, 10, 778);SEQ. ID NO: 82 QPQMSHM (Sun et al. Polymers 2018, 10, 778);SEQ. ID NO: 83 TNIESLK (Sun et al. Polymers 2018, 10, 778);SEQ. ID NO: 84 NCHKCWN (Sun et al. Polymers 2018, 10, 778);SEQ. ID NO: 85 SHLSKNF (Sun et al. Polymers 2018, 10, 778).

Immunoglobulin E (IgE)

In some embodiments, the Target Protein is human immunoglobulin E (IgE).IgE is a type of immunoglobulin that plays an essential role in type Ihypersensitivity, which can manifest into various allergic diseases,such as allergic asthma, most types of sinusitis, allergic rhinitis,food allergies, and specific types of chronic urticaria and atopicdermatitis. IgE also plays a pivotal role in responses to allergens,such as: anaphylactic drugs, bee stings, and antigen preparations usedin desensitization immunotherapy.

The Protein Data Bank website provides the crystal structure of IgEsearchable by 1F2Q (Garman, S. C., Kinet, J. P., Jardetzky, T. S., Cell,1998, 95: 951-961); as well as the crystal structure of IgE bound tovarious compounds searchable by 1F6A (Garman, S.C., et al., Nature,2000, 406 259-266); 1RPQ (Stamos, J., et al., Structure, 2004, 121289-1301); 2Y7Q (Holdom, M. D., et al., Nat. Struct. Mol. Biol., 2011,18 571); and 4GRG (Kim, B., et al., Nature, 2012, 491: 613-617).

Additionally, Wan et al., provides insight into the crystal structure ofIgE Fc, revealing an asymmetrically bent conformation (Wan et al., Nat.Immunol., 2002, 3(7), 681-6); and Dhaliwal et al, provides insight intothe crystal structure of IgE bound to its B-cell receptor CD23 reveals amechanism of reciprocal allosteric inhibition with high affinityreceptor FcεRI (Dhaliwal, B., et al., Proc Natl Acad Sci USA., 2012,109(31), 12686-91).

Additional Immunoglobin Targeting Ligands

Additional, non-limiting examples of Extracellular Targeting Ligandsinclude:

wherein X_(M) is —(CH₂)₀₋₆, —O—(CH₂)₀₋₆, S—(CH₂)₀₋₆, NR_(M)—(CH₂)₀₋₆,C(O)—(CH₂)₀₋₆, a PEG group containing from 1 to 8, preferably 1-4ethylene glycol residues, or a —C(O)(CH₂)₀₋₆NR_(M) group; R_(M) is H ora C₁-C₃ alkyl group which is optionally substituted with one or twohydroxyl groups, where 0-6 is preferably 1, 2, 3, or 4, more preferably1.

Additional, non-limiting examples of Extracellular Targeting Ligandsinclude:

wherein DNP is a 2,4-dinitrophenyl group; or a group according to thechemical structure:

wherein Y′ is H or NO₂ (preferably H);X¹⁰¹ is O, CH₂, S, NR¹⁰¹, S(O), S(O)₂, —S(O)₂O, —OS(O)₂, or OS(O)₂O; andR¹⁰¹ is H, a C₁-C₃ alkyl group, or a —C(O)(C₁-C₃ alkyl) group.

Additional, non-limiting examples of Extracellular Targeting Ligandsinclude:

wherein X¹⁰² is CH, O, N—R¹⁰¹, or S, preferably O;R¹⁰¹ is H or C₁-C₃ alkyl; andZ is a bond, a monosaccharide, disaccharide, oligosaccharide, morepreferably a sugar group selected from the monosaccharides, includingaldoses and ketoses, and disaccharides, including those disaccharidesdescribed herein. Monosaccharide aldoses include monosaccharides such asaldotriose (D-glyceraldehdye, among others), aldotetroses (D-erythroseand D-Threose, among others), aldopentoses, (D-ribose, D-arabinose,D-xylose, D-lyxose, among others), aldohexoses (D-allose, D-altrose,D-Glucose, D-Mannose, D-gulose, D-idose, D-galactose and D-Talose, amongothers), and the monosaccharide ketoses include monosaccharides such asketotriose (dihydroxyacetone, among others), ketotetrose (D-erythrulose,among others), ketopentose (D-ribuulose and D-xylulose, among others),ketohexoses (D-Psicone, D-Fructose, D-Sorbose, D-Tagatose, amongothers), aminosugars, including galactoseamine, sialic acid,N-acetylglucosamine, among others and sulfosugars, includingsulfoquinovose, among others.

Exemplary disaccharides which find use in the present invention includesucrose (which may have the glucose optionally N-acetylated), lactose(which may have the galactose and/or the glucose optionallyN-acetylated), maltose (which may have one or both of the glucoseresidues optionally N-acetylated), trehalose; (which may have one orboth of the glucose residues optionally N-acetylated), cellobiose (whichmay have one or both of the glucose residues optionally N-acetylated),kojibiose (which may have one or both of the glucose residues optionallyN-acetylated), nigerose (which may have one or both of the glucoseresidues optionally N-acetylated), isomaltose (which may have one orboth of the glucose residues optionally N-acetylated), b,b-trehalose(which may have one or both of the glucose residues optionallyN-acetylated), sophorose (which may have one or both of the glucoseresidues optionally N-acetylated), laminaribiose (which may have one orboth of the glucose residues optionally N-acetylated), gentiobiose(which may have one or both of the glucose residues optionallyN-acetylated), turanose (which may have the glucose residue optionallyN-acetylated), maltulose (which may have the glucose residue optionallyN-acetylated), palatinose (which may have the glucose residue optionallyN-acetylated), gentiobiluose (which may have the glucose residueoptionally N-acetylated), mannobiose, melibiose (which may have theglucose residue and/or the galactose residue optionally N-acetylated),melibiulose (which may have the galactose residue optionallyN-acetylated), rutinose, (which may have the glucose residue optionallyN-acetylated), rutinulose and xylobiose, among others.

TNF-α

In some embodiments, the Target Protein is human TNF-α (UniProtKB—P01375(TNFA_HUMAN)). TNF-α is a pro-inflammatory cytokine active in the bodilyimmune response and serious inflammatory diseases. TNF-α has beenimplicated in a number of disorders, including but not limited torheumatoid arthritis, inflammatory bowel disease, graft-vs-host disease,ankylosing spondylitis, psoriasis, hidradenitis suppurativa, refractoryasthma, systemic lupis erthyematosus, diabetes, and the induction ofcachexia.

The Protein Data Bank website provides the crystal structure of TNF-αsearchable by 6RMJ (Valentinis, B., et al., Int. J. Mol. Sci., 2019,20); 5UUI (Carrington et al., Biophys J., 2017, 113 371-380); 600Y, 600Zand 60PO (O'Connell, J., et al., Nat. Commun., 2019, 10 5795-5795); and5TSW (Cha, S. S., J Biol Chem., 1998, 273 2153-2160); as well as thecrystal structure of TNF-α bound to various compounds searchable by 5YOY(Ono et al., Protein Sci., 2018, 27 1038-1046); 2AZ5 (He., M. M., etal., Science, 2005, 310: 1022-1025); 5WUX (Lee, J. U., Int J Mol Sci.,2017, 18); 5MU8 (Blevitt et al., J Med Chem., 2017, 60 3511-3517); 4Y60(Feldman J. L., et al., Biochemistry, 2015, 54 3037-3050); 3WD5 (Hu, S.,et al., J Biol Chem, 2013, 288 27059-27067); and 4G3Y (Liang, S. Y., JBiol Chem., 2013, 288 13799-13807).

Representative TNF-α Targeting Ligands are provided in FIG. 1 .Additional TNF-α Targeting Ligands can be found in, for example, U.S.Pat. No. 8,541,572; J Chem Inf Model. 2017 May 22; 57(5): 1101-1111;each of which is incorporated by reference herein.

IL-1

In some embodiments, the Target Protein is human interleukin-1 (IL-1)(UniProtKB-P01584 (IL1B HUMAN)). IL-1 is a potent proinflammatorycytokine. Initially discovered as the major endogenous pyrogen, inducesprostaglandin synthesis, neutrophil influx and activation, T-cellactivation and cytokine production, B-cell activation and antibodyproduction, and fibroblast proliferation and collagen production. IL-1promotes Th17 differentiation of T-cells, and Synergizes withIL12/interleukin-12 to induce IFNG synthesis from T-helper 1 (Th1)cells. IL-1 has been implicated in a number of auto-inflammatory andautoimmune disorders, including, but not limited to, Blau syndrome,cryopyrin-associated periodic syndromes, familial Mediterranean fever,Majeed syndrome; mevalonate kinase deficiency syndrome, pyogenicarthritis-pyoderma gangrenosum-acne syndrome, tumor necrosis factorreceptor-associated periodic syndrome, Behget's Disease, Sjogren'sSyndrome, gout and chondrocalcinosis, periodic fever, aphthousstomatitis, pharyngitis, and cervical adenitis (or PFAPA) syndrome,rheumatoid arthritis, Type 2 diabetes mellitus, acute pericarditis,Chronic interstitial lung diseases (ILDs), Still's Disease,

The Protein Data Bank website provides the crystal structure of IL-1searchable by 9ILB (Yu, B., et al., Proc Natl Acad Sci USA, 1999, 96103-108); 1I1B (Finzel, B. C., et al., J Mol Biol., 1989, 209 779-791);and 3040 (Wang et al., Nat.Immunol., 2010, 11: 905-911); as well as thecrystal structure of IL-1 bound to various compounds searchable by 4G6J(Blech, M., et al., J Mol Biol., 2013, 425 94-111); 5BVP (Rondeau e al.,MAbs, 2015, 7 1151-1160); and 3LTQ (Barthelmes, K., et al., J Am Chem.Soc., 2011, 133 808-819). Additionally, Guy et al., provides insightinto the crystal structure of a small antagonist peptide bound tointerleukin-1 receptor type 1 (Guy et al., The Journal of BiologicalChemistry, 2000, 275, 36927-36933).

Potential IL-1 direct or indirect inhibitors are described in FIG. 1 .Additional IL-1 Targeting Ligands can be found in, for example, U.S.Pat. No. 9,694,015, each of which is incorporated herein by reference.Additional binding ligands include rilanocept or a binding fragmentthereof (J Rheumatol. 2012; 39:720-727 (2012); and Canakinurnab, or abinding fragment thereof (J Rheumatol. 2004;31:I 103-111 1).

IL-2

In some embodiments, the Target Protein is human interleukin-2 (IL-2)(UniProtKB-P60568 (IL2_HUMAN)). IL-2 is a potent pro-inflammatorycytokine. IL-2 has been implicated in host versus graft rejection andother autoimmune disorders.

The Protein Data Bank website provides the crystal structure of IL-2searchable by 1M4C and 1M47 (Arkin, M. R., et al.,Proc.Natl.Acad.Sci.USA, 2003, 100: 1603-1608); as well as the crystalstructure of IL-2 bound to various compounds searchable by 4NEJ and 4NEM(Brenke, R., et al.); 1QVN (Thanos, C. D., et al., Proc Natl Acad SciUSA, 2006, 103 15422-15427); 1PW6 and 1PY2 (Thanos, C. D., et al., J AmChem Soc., 2003, 125 15280-15281); 1NBP (Hyde, J., et al., Biochemistry,2003, 42 6475-6483); and 1M48, 1M49, 1M4A, 1M4B, and 1M4C (Arkin, M. R.,et al., Proc Natl Acad Sci USA, 2003, 100 1603-1608). Additionally,Stauber, D. J., et al, provides insight into the crystal structure ofthe IL-2 signaling complex: paradigm for a heterotrimeric cytokinereceptor (Stauber, D. J., et al., PNAS, 2006, 103(8), 2788-2793).

Representative IL-2 Targeting Ligands are provided in FIG. 1 .Additional IL-2 Targeting Ligands can be found in, for example, U.S.Pat. Nos. 8,802,721; 9,682,976, 9,708,268; Eur J Med Chem 83: 294-306(2014), J Med Chem 60: 6249-6272 (2017); Nature 450: 1001-1009 (2007);each of which is incorporated by reference herein.

IL-6

In some embodiments, the Target Protein is human inteleukin-6 (IL-6)(UniProtKB-P05231 (IL6_HUMAN)). IL-6 is a cytokine with a wide varietyof biological functions. It is a potent inducer of the acute phaseresponse and plays an essential role in the final differentiation ofB-cells into Ig-secreting cells. It is also involved in lymphocyte andmonocyte differentiation. It also acts on B-cells, T-cells, hepatocytes,hematopoietic progenitor cells and cells of the CNS, and is required forthe generation of T(H)₁₇ cells. IL-6 has been implicated in a number ofinflammatory diseases and cancers, including, but not limited to,Castleman's disease, metastatic castration-associated prostate cancer,renal cell carcinoma, large-cell lung carcinoma, ovarian cancer,rheumatoid arthritis, asthma.

The Protein Data Bank website provides the crystal structure of IL-6searchable by 1P9M (Boulanger, M. J., et al., Science, 2003, 300:2101-2104); IALU (Somers et al., EMBO J., 1997, 16, 989-997); 1IL6 and2IL6 (Xu, G. Y., et al., J Mol Biol., 1997, 268 468-481) and 1N26(Varghese et al., Proc Natl Acad Sci USA., 2002, 99 15959-15964); aswell as the crystal structure of IL-6 bound to various compoundssearchable by 4CNI (Shaw, S., et al., Mabs, 2014, 6: 773); and 4NI7 and4NI9 (Gelinas et al., J Biol Chem. 2014, 289(12), 8720-8734).Additionally, Gelinas et al., provides insight into the crystalstructure of interleukin-6 in complex with a modified nucleic acidligand (Gelinas, A. D., et al., J Biol Chem. 2014, 289(12), 8720-8734);and Somers et al., provides insight into the crystal structure ofinterleukin 6: implications for a novel mode of receptor dimerizationand signaling.

Potential IL-6 direct or indirect inhibitors are provided in FIG. 1 .Additional potential IL-6 direct or indirect inhibitors can be found in,for example, U.S. Pat. No. 8,901,310; U.S. patent Ser. No. 10/189,796;U.S. Pat. No. 9,694,015; each incorporated herein by reference. Inanother embodiment the IL-6 Extracellular Targeting Ligand is AvimarC326or a binding fragment thereof which is described in Nat Biotechnol 23,1556-1561 (2005).

IFN-γ

In some embodiments, the Target Protein is human interferon-γ (IFN-γ)(UniProtKB-Q14609 (Q14609_HUMAN)). IFN-γ is a immunoregulatory cytokine.IFN-γ has been implicated in a number of autoimmune disorders,including, but not limited to rheumatoid arthritis, multiple sclerosis(MS), corneal transplant rejection, and various autoimmune skin diseasessuch as psoriasis, alopecia areata, vitiligo, acne vulgaris, and others.

The Protein Data Bank website provides the crystal structure of IFN-γsearchable by 1HIG (Ealick, S. E., et al., Science 252, 1991, 698-702);as well as the crystal structure of IFN-γ bound to various compoundssearchable by 6E3K and 6E3L (Mendoza, J. L., et al., Nature, 2019, 56756-60). Additionally, Randal et al., provides insight into the structureand activity of a monomeric interferon-γ: α-chain receptor signalingcomplex (Randal, M., et al., Structure, 2001, 9(2), 155-163).

Representative IFN-γ Targeting Ligands are described in FIG. 1 .Additional IFN-7 Targeting Ligands can be found in, for example, J MedChem 57: 4511-20 (2014); which is incorporated by reference herein.

Vascular Epithelial Growth Factor (VEGF)

In some embodiments, the Target Protein is human vascular epithelialgrowth factor (VEGF) (UniProtKB—P15692 (VEGFA_HUMAN)). VEGF is a growthfactor active in angiogenesis, vasculogenesis, and endothelial cellgrowth. VEGF induces endothelial cell proliferation, promotes cellmigration, inhibits apoptosis and induces permeabilization of bloodvessels. VEGF has been implicated in the vascularization andangiogenesis of tumors.

The Protein Data Bank website provides the crystal structure of VEGFsearchable by 3QTK (Mandal, K., et al., Angew Chem Int Ed Engl., 2011,50 8029-8033); and 4KZN (Shen et al.); as well as the crystal structureof VEGF bound to various compounds searchable by 504E (Lobner, E., etal., MAbs, 2017, 9 1088-1104); 4QAF (Giese, T., et al.,); 5DN2 (Tsai, Y.C. I., et al., FEBS, 2017, J 283 1921-1934); 4GLS (Mandal, K., et al.,Proc Natl Acad Sci USA, 2012, 109 14779-14784); and 1KMX (Stauffer, M.E. et al., J Biomol NMR, 2002, 23 57-61). Additionally, Mueller, Y. A.,et al, provides insight into the Crystal structure and functionalmapping of the kinase domain receptor binding site of VEGF (Mueller, Y.A., et al., Proc Natl Acad Sci USA., 1997 Jul 8; 94(14): 7192-7197).

Representative VEGF Targeting Ligands are provided in FIG. 1 .Additional VEGF Targeting Ligands include, but are not limited to, (allcited referenced incorporated herein by reference) the peptideVEPNCDIHVMWEWECFERL-NH₂ (Biochemistry 1998, 37, 17754-177764).Additional VEGF Targeting Ligands are provided in, for example, J MedChem 57: 3011-29 (2014), U.S. Pat. Nos. 9,884,843, 9,446,026, J Med Chem53: 1686-99 (2010), J Med Chem 48: 8229-36 (2005), J Nat Prod 76: 29-35(2013), each of which is incorporated herein by reference.

Transforming Growth Factor-β1 (TGF-β1)

In some embodiments, the Target Protein is human transforming growthfactor-01 (TGF-β1) (UniProtKB—P01137 (TGFB1_HUMAN)). TGF-β1 is amultifunctional protein that regulates the growth and differentiation ofvarious cell types and is involved in various processes, such as normaldevelopment, immune function, microglia function and responses toneurodegeneration. TGF-β1 can promote either T-helper 17 cells (Th17) orregulatory T-cells (Treg) lineage differentiation in aconcentration-dependent manner. TGF-β1 expression in the tumormicroenvironment has been associated with a poor prognosis, and isimplicated in TGF-β1 mediated tumor suppression via T-cell exclusion.TGF-β1 expression has also been implicated in hematological malignanciesand fibrosis.

The Protein Data Bank website provides the crystal structure of TGF-β1searchable by 5E8S, 5E8T, and 5E8U (Tebben, A. J., et al., ActaCrystallogr D Struct Biol., 2016, 72 658-674); 2L5S (Zuniga, J. E., etal, J Mol Biol., 2011, 412 601-618); and 2PJY (Groppe, J., et al., MolCell, 2008, 29 157-168); as well as the crystal structure of TGF-β1bound to various compounds searchable by 5QIK, 5QIL and 5QIM, (Zhang,Y., et al., ACS Med Chem Lett., 2018, 9 1117-1122); 6B8Y (Harikrishnan,L. S., et al., Bioorg Med Chem., 2018, 26 1026-1034); 5E8W, 5E8X, 5E8Z,and 5E90 (Tebben, A. J., et al., Acta Crystallogr D Struct Biol., 2016,72 658-674); 3TZM (Ogunjimi, A. A. et al., Cell Signal, 2012, 24476-483); 2X70 (Roth, G. J., et al., J Med Chem., 2010, 53 7287); 3KCF(Guckian, K., et al., Bioorg Med Chem Lett., 2010, 20 326-329); 3FAA(Bonafoux, D., et al., Bioorg Med Chem Lett., 2009, 19 912-916); 1VJY(Gellibert, F, J., et al., J Med Chem., 2004 47 4494-4506); and 1PY5(Sawyer, J. S., et al., Bioorg Med Chem Lett., 2004, 14 3581-3584).Additionally, Hinck et al., provides insight into the structural studiesof the TGF-βs and their receptors and further insight into evolution ofthe TGF-β superfamily (Hinck, A., FEBS, 2012, 586(14), 1860-1870).

Representative TGF-β1 Targeting Ligands are provided in FIG. 1 . In someembodiments, the TGF-β1 Targeting Ligand is the peptide KRFK peptide (J.Biol. Chem. Vol. 274 (No. 19) pp. 13586-13593 (1999)(incorporated hereinby reference). Additional TGF-β1 Targeting Ligands are provided in, forexample, Bioorg Med Chem Lett 21: 5642-5 (2011), which is incorporatedherein by reference.

Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK-9)

In some embodiments, the Target Protein is human proprotein convertasesubtilisin/kexin type 9 (PCSK-9) (UniProtKB—Q8NBP7 (PCSK9_HUMAN)).PCSK-9 is a crucial player in the regulation of plasma cholesterolhomeostasis. PCSK-9 binds to low-density lipid receptor family members:low density lipoprotein receptor (LDLR), very low-density lipoproteinreceptor (VLDLR), apolipoprotein E receptor (LRP1/APOER) andapolipoprotein receptor 2 (LRP8/APOER2), and promotes their degradationin intracellular acidic compartments. It acts via a non-proteolyticmechanism to enhance the degradation of the hepatic LDLR through aclathrin LDLRAPl/ARH-mediated pathway, and may prevent the recycling ofLDLR from endosomes to the cell surface or direct it to lysosomes fordegradation. PCSK-9 has been implicated in high blood cholesterol andthe development of cardiovascular disease.

The Protein Data Bank website provides the crystal structure of PCSK-9searchable by 2P4E (Cunningham, D., et al., Nat Struct Mol Biol., 2007,14 413-419); as well as the crystal structure of PCSK-9 bound to variouscompounds searchable by 3BPS (Kwon, H. J., et al., Proc Natl Acad SciUSA, 2008, 105 1820-1825); 6U26, 6U2N, 6U2P, 6U36, 6U38, and 6U3X(Petrilli, W. L., et al., Cell Chem Biol., 2019, 27 32-40.e3); 50CA(Gustafsen, C., et al., Nat Commun., 2017, 8 503-503); 4NE9 (Schroeder,C. I., et al., Chem Biol., 2014, 21 284-294); 40V6 (Mitchell, T., etal., J Pharmacol Exp Ther., 2014, 350 412-424); and 4NMX (Zhang, Y., etal., J Biol Chem., 2014, 289 942-955). Additionally, Piper et al.,provides insight into the crystal structure of PCSK9 (Piper, D. E., etal., Structure, 2007, 15(5), 545-52).

Representative PCSK-9 Targeting Ligands are provided in FIG. 1 . In someembodiments, the PCSK-9 Targeting Ligand is the peptide TVFTSWEEYLDWV(J. Bio. Chem. 2014 January; 289(2):942-955, incorporated herein byreference). Additional PCSK-9 Targeting Ligands are provided in, forexample, U.S. Pat. No. 9,227,956, J Biol Chem 289: 942-55 (2014), eachof which is incorporated by reference herein.

IL-21

In some embodiments, the Target Protein is human interleukin-21 (IL-21)(UniProtKB-Q9HBE4 (IL21_HUMAN)). IL-21 is an immunoregulatory cytokine.IL-21 has been implicated in a number of autoimmune disorders, includingSjogren's syndrome, systemic lupus erythematosus, type 1 diabetes,multiple sclerosis, rheumatoid arthritis, and inflammatory boweldisease.

The Protein Data Bank website provides the crystal structure of IL-21searchable by 2OQP (Bondensgaard, K., et al., J Biol Chem., 2007, 28223326-23336); and 4NZD (Hamming et al.); as well as the crystalstructure of IL-21 bound to various compounds searchable by 3TGX(Hamming, 0. J., et al., J Biol Chem., 2012, 287(12), 9454-9460).

Representative IL-21 Targeting Ligands are described in FIG. 1 .Additional IL-21 Targeting Ligands can be found in, for example, U.S.Pat. No. 9,701,663, which is incorporated herein by reference.

IL-22

In some embodiments, the Target Protein is human interleukin-22 (IL-22)(UniProtKB-Q9GZX6 (IL22_HUMAN)). IL-22 is a member of IL-10 familycytokines that is produced by many different types of lymphocytesincluding both those of the innate and adaptive immune system. IL-22 hasbeen implicated in a number of autoimmune disorders, including, but notlimited to, graft versus host disease (GVHD), psoriasis, rheumatoidarthritis, atopic dermatitis, and asthma.

The Protein Data Bank website provides the crystal structure of IL-22searchable by 1M4R (Nagem, R. A. P., et al., Structure, 2002, 101051-1062); as well as the crystal structure of IL-22 bound to variouscompounds searchable by 3DGC (Jones, B. C. et al., Structure, 2008, 161333-1344).

Representative IL-22 Targeting Ligands are described in FIG. 1 .Additional IL-22 Targeting Ligands can be found in, for example, U.S.Pat. No. 9,701,663, which is incorporated herein by reference.

IL-10

In some embodiments, the Target Protein is human interleukin-10 (IL-10)(UniProtKB-P22301 (IL10_HUMAN)). IL-10 is an inflammatory cytokine.IL-10 has been implicated in tumor survival and protection againstcytotoxic chemotherapeutic drugs.

The Protein Data Bank website provides the crystal structure of IL-10searchable by 2ILK (Zdanov, A et al., Protein Sci., 1996, 5 1955-1962);1ILK (Zdanov, A. et al., Structure, 1995, 3 591-601); 2H24 (Yoon, S. I.,et al., J Biol Chem., 2006, 281 35088-35096) and 3LQM (Yoon, S. I., etal., Structure, 2010, 18 638-648). Additionally, Zdanov, A., et al,provides insight into crystal structure of IL-10 (Zdanov A., CurrentPharmaceutical design, 2004, 10, 3873-3884).

Representative IL-10 Targeting Ligands are provided in FIG. 1 .Additional IL-10 Targeting Ligands can be found, for example, in ACSChem Biol 11: 2105-11 (2016), which is incorporated herein by reference.

IL-5

In some embodiments, the Target Protein is human interleukin-5 (IL-5)(UniProtKB-P05113 (IL5 HUMAN)). IL-5 is a cytokine that regulateseosinophil maturation, recruitment, and survival. IL-5 has beenimplicated in a number of allergic disorders, including, but not limitedto, asthma, nasal polyposis, atopic dermatitis, eosinophilicesophagitis, hypereosinophilic syndrome, and Churg-Strauss syndrome.

The Protein Data Bank website provides the crystal structure of IL-5searchable by 1HUL (Milburn, M. V., Nature, 1993, 363, 172-176) and 3VA2(Kusano et al., Protein Sci., 2012, 21(6), 850-864); as well as thecrystal structure of IL-5 bound to various compounds searchable by 1OBXand 1OBZ (Kang, B. S., et al., Structure, 2003, 11, 845).

Representative IL-5 Targeting Ligands are provided in FIG. 1 .Additional IL-5 Targeting Ligands can be found, for example, in BioorgMed Chem 18: 4441-5 (2010); Bioorg Med Chem 18: 4625-9 (2011); BioorgMed Chem 21: 2543-50 (2013); Eur J Med Chem 59: 31-8 (2013); Bioorg MedChem 23: 2498-504 (2015); Bioorg Med Chem 20: 5757-62 (2012); each ofwhich is incorporated by reference herein.

IL8

In some embodiments, the Target Protein is human interleukin-8 (IL-8)(UniProtKB-P10145 (IL8_HUMAN)). IL-8 is a chemotactic factor thatattracts neutrophils, basophils, and T-cells, but not monocytes. It isalso involved in neutrophil activation. It is released from several celltypes in response to an inflammatory stimulus. IL-8 has been implicatedin the promotion of tumor progression, immune escape,epithelial-mesenchymal transition, and recruitment of myeloid-derivedsuppressor cells. Studies have demonstrated that high serum IL-8 levelscorrelate with poor prognosis in many malignant tumors. Preclinicalstudies have shown that IL-8 blockade may reduce mesenchymal features intumor cells, making them less resistant to treatment.

The Protein Data Bank website provides the crystal structure of IL-8searchable by 3IL8 (Baldwin, E. T., et al., Proc Natl Acad Sci USA,1991, 88, 502-506); and 1IL8 and 2IL8 (Clore, G. M., et al.,Biochemistry, 1990, 29, 1689-1696); as well as the crystal structure ofIL-8 bound to various compounds searchable by 1ILP and 1ILQ (Skelton, N,J., et al., Structure, 1999, 7, 157-168); and 1ROD (Sticht, H., et al.,Eur J Biochem., 1996, 235, 26-35); 4XDX (Ostrov et al.,) and 5WDZ(Beckamp, S., J Biomol NMR, 2017, 69, 111-121).

Representative IL-8 Targeting Ligands are provided in FIG. 1 .Additional IL-8 Targeting Ligands can be found in, for example, BioorgMed Chem Lett 19: 4026-30 (2009), which is incorporated by referenceherein.

Cholinesterase

In some embodiments, the Target Protein is human cholinesterase(UniProtKB—P06276 (CHLE_HUMAN)). Cholinesterase contributes to theinactivation of the neurotransmitter acetylcholine. Inhibition ofcholinesterase results in increased levels of acetylcholine in thesynaptic cleft (the space between two nerve endings). The main use ofcholinesterase inhibitors is for the treatment of dementia in patientswith Alzheimer's disease. People with Alzheimer's disease have reducedlevels of acetylcholine in the brain. Cholinesterase inhibitors havebeen shown to have an effect on dementia symptoms such as cognition.

The Protein Data Bank website provides the crystal structure ofcholinesterase searchable by 1POI and 1POQ (Nicolet, Y., et al., J BiolChem., 2003, 278, 41141-41147); as well as the crystal structure ofcholinesterase bound to various compounds searchable by 1POM and 1POP(Nicolet, Y., et al., J Biol Chem., 2003, 278, 41141-41147); 2J4C(Frasco, M. F., et al., FEBS J., 2007, 274 1849); 4BDT, 4BDS (Nachon,F., et al., Biochem J, 2013, 453, 393-399); 1GQR and 1GQS (Bar-on, P.,et al., Biochemistry, 2002, 41, 3555); 3DJY and 3DKK (Carletti, E., etal., J Am Chem Soc., 2008, 130, 16011-16020); 4AXB, 4B00, 4BOP, and 4BBZ(Wandhammer, M., et al., Chem Biol Interact., 2013, 203, 19); 1DX6(Greenblatt, H. M., et al., FEBS Lett., 1999, 463 321); 1GPK and 1GPN(Dvir, H., et al., Biochemistry, 2002, 41, 10810); 6CQY (Bester, S. M.,et al., Chem Res Toxicol., 2018, 31, 1405-1417); 1XLV and 1XLW (Nachon,F., et al., Biochemistry, 2005, 44, 1154-1162); 2Y1K (Carletti, E., etal., Chem Res Toxicol., 2011, 24, 797); and 2WIG, 2WIJ, 2WIK, 2WIL, and2WSL (Carletti, E., et al., Biochem J., 2009, 421, 97-106).Additionally, Ahmad et al., provides insight into the isolation, crystalstructure determination and cholinesterase inhibitory potential ofisotalatizidine hydrate from delphinium denudatum (Ahmad H., et al.,Journal Pharmaceutical Biology, 2016, 55(1), 680-686).

Representative cholinesterase Targeting Ligands are provided in FIG. 1 .Additional Targeting Ligands can be found in, for example, ACS Med ChemLett 4: 1178-82 (2013); J Med Chem 49: 3421-5 (2006); Eur J Med Chem 55:23-31 (2012); J Med Chem 51: 3154-70 (2008); J Med Chem 46: 1-4 (2002);Eur J Med Chem 126: 652-668 (2017); Biochemistry 52: 7486-99 (2013);Bioorg Med Chem 23: 1321-40 (2015); which are each incorporated hereinby reference.

C-C Motif Chemokine Ligand 2 (CCL2)

Grygiel et al., provides insight into the synthesis by native chemicalligation and crystal structure of human CCL2 (Grygiel, T. L., et al.,Biopolymers, 2010, 94(3), 350-9).

In some embodiments, the Target Protein is human C-C motif chemokineligand 2 (CCL2) (UniProtKB—P13500 (CCL2_HUMAN)). CCL2 acts as a ligandfor C-C chemokine receptor CCR2. CCL2 signals through binding andactivation of CCR2 and induces a strong chemotactic response andmobilization of intracellular calcium ions. CCL2 exhibits a chemotacticactivity for monocytes and basophils but not neutrophils or eosinophils.

CCL2 has been implicated in the recruitment of monocytes into thearterial wall during the disease process of atherosclerosis.

Representative CCL2 Targeting Ligands are provided in FIG. 1 .Additional CCL2 Targeting Ligands can be found in, for example, J MedChem 56: 7706-14 (2013), which is incorporated herein by reference.

Carboxypeptidase B2

In some embodiments, the Target Protein is human carboxypeptidase B2(UniProtKB-Q96IY4 (CBPB2_HUMAN)). Carboxypeptidase B2, also known asthrombin activatable fibrinolysis inhibitor (TAFIa), cleaves C-terminalarginine or lysine residues from biologically active peptides such askinins or anaphylatoxins in the circulation thereby regulating theiractivities. It down-regulates fibrinolysis by removing C-terminal lysineresidues from fibrin that has already been partially degraded byplasmin. Carboxypeptidase B2 has been implicated and targeted to inhibitthrombosis.

The Protein Data Bank website provides the crystal structure ofcarboxypeptidase B2 (also known as thrombin-activatable fibrinolysisinhibitor (TAFI)) searchable by 3D66 (Marx, P. F., et al., Blood, 2008,112, 2803-2809); 3DGV (Anand, K., et al., JBC, 2008, 283, 29416-29423);and 1KWM (Barbosa Pereira, P. J., et al., J Mol Biol., 2002, 321,537-547); as well as the crystal structure of TAFI bound to variouscompounds searchable by 3D67 (Marx, P. F., et al., Blood, 2008, 112,2803-2809); 5HVF, 5HVG, 5HVH (Zhou, X., et al., J Thromb Haemost., 2016,14, 1629-1638); and 3LMS (Sanglas, L., et al., J Thromb Haemost., 2010,8, 1056-1065). Additionally, Schreuder et al., provides insight into theinteraction of TAFI and anabaenopeptin, a highly potent inhibitor ofTAFI (Schreuder, H., et al., Sci Rep., 2016, 6, 32958).

Representative carboxypeptidase B2 Targeting Ligands are provided inFIG. 1 . Additional carboxypeptidase B2 Targeting Ligands can be foundin, for example, Bioorg Med Chem Lett 20: 92-6 (2010), J Med Chem 50:6095-103 (2007), Bioorg Med Chem Lett 14: 2141-5 (2004), J Med Chem 58:4839-44 (2015), J Med Chem 55: 7696-705 (2012), J Med Chem 59: 9567-9573(2016), Bioorg Med Chem Lett 17: 1349-54 (2007), U.S. Pat. Nos.9,662,310, 8,609,710, 9,688,645, J Med Chem 46: 5294-7 (2003), each ofwhich is incorporated herein by reference.

Neutrophil Elastase

In some embodiments, the Target Protein is human neutrophil elastase(UniProtKB-P08246 (ELNE_HUMAN)). Neutrophil elastase modifies thefunctions of natural killer cells, monocytes and granulocytes. InhibitsC5a-dependent neutrophil enzyme release and chemotaxis.

Neutrophil elastase has been implicated in a number of disorders,including lung disease, chronic obstructive pulmonary disease,pneumonia, respiratory distress, and acute lung injury (ALI), and cysticfibrosis, as well as chronic kidney disease.

The Protein Data Bank website provides the crystal structure of humanneutrophil elastase bound to various compounds searchable by 3Q76 and3Q77 (Hansen, G., et al., J.Mol.Biol., 2011, 409, 681-691); 5ABW (VonNussbaum, et al., Bioorg Med Chem Lett., 2015, 25, 4370-4381); 1BOF(Cregge, R. J., et al., J Med Chem., 1998, 41, 2461-2480); 1H1B(Macdonald, S. J. F., et al., J Med Chem., 2002, 45, 3878); 2Z7F(Koizumi, M., et al., J Synchrotron Radiat., 2008, 15 308-311); 5A09,5AOA, 5AOB, and 5AOC (Von Nussbaum, F., et al., Chem Med Chem., 2015,10, 1163-1173); 5A8X, 5A8Y and 5A8Z (Von Nussbaum, F., et al.,ChemMedChem., 2016, 11, 199-206); 1HNE (Navia, M. A., et al., Proc NatlAcad Sci USA, 1989, 86, 7-11); 6F5M (Hochscherf, J., et al., ActaCrystallogr F Struct Biol Commun., 2018, 74, 480-489); and 4WVP(Lechtenberg, B. C., et al., ACS Chem Biol., 2015, 10, 945-951).

Representative neutrophil elastase Targeting Ligands are provided inFIG. 1 . Additional neutrophil elastase Targeting Ligands can be foundin, for example, J Med Chem 53: 241-53 (2010), J Med Chem 38: 739-44(1995), J Med Chem 37: 2623-6 (1994), J Med Chem 38: 4687-92 (1995), JMed Chem 45: 3878-90 (2002), Bioorg Med Chem Lett 5: 105-109 (1995),Bioorg Med Chem Lett 11: 243-6 (2001), J Med Chem 40: 1906-18 (1997),Bioorg Med Chem Lett 25: 4370-81 (2015), U.S. Pat. Nos. 8,569,314,9,174,997, 9,290,457, each of which is incorporated herein by reference.

Factor Xa

In some embodiments, the Target Protein is human Factor Xa(UniProtKB—P00742 (FA10_HUMAN)). Factor Xa is a vitamin K-dependentglycoprotein that converts prothrombin to thrombin in the presence offactor Va, calcium and phospholipid during blood clotting.

Factor X has been implicated in the development of deep vein thrombosisand acute pulmonary embolism, and the risk of stroke and embolism inpeople with nonvalvular atrial fibrillation.

The Protein Data Bank website provides the crystal structure of FactorXa bound to various compounds searchable by 1G2L and 1G2M (Nar, H., etal., Structure, 2001, 9, 29-38); 2PR3 (Nan huis, C. A., et al., ChemBiol Drug Des., 2007, 69, 444-450); 2UWP (Young, R. J., et al., BioorgMed Chem Lett., 2007, 17, 2927); 2VVC, 2VVV, 2VVU, 2VWL, 2VWM, 2VWN and2VWO (Zbinden, K. G., et al., Eur J Med Chem., 2009, 44, 2787); 4Y6D,4Y71, 4Y7A, 4Y7B, 4zh8, 4ZHA (Convery, M. A. et al.); 4Y76, 4Y79, 2J94and 2J95 (Chan, C., et al., J Med Chem., 2007, 50 1546-1557); 1FAX(Brandstetter, H., et al., J Biol Chem., 1996, 271, 29988-29992); 2JKH(Salonen, L. M., et al., Angew Chem Int Ed Engl., 2009, 48, 811); 2PHB(Kohrt, J. T., et al., Chem Biol Drug Des., 2007, 70, 100-112); 2W26(Roehrig, S., et al., J Med Chem., 2005, 48, 5900); 2Y5F, 2Y5G and 2Y5H(Salonen, L. M., et al., Chemistry, 2012, 18, 213); 3Q3K (Yoshikawa, K.,et al., Bioorg Med Chem Lett., 2011, 21, 2133-2140); 2BMG (Matter, K.,et al., J Med Chem., 2005, 48, 3290); 2BOH, 2BQ6 2BQ7, and 2BQW (Nazare,M., et al., J Med Chem., 2005, 48, 4511); 2CJI (Watson, N. S., et al,Bioorg Med Chem Lett., 2006, 16, 3784); 2J2U, 2J34, 2J38, 2J41 (Senger,S., et al., Bioorg Med Chem Lett., 2006, 16 5731); 3IIT (Yoshikawa, K.,et al.,Bioorg Med Chem., 2009, 17 8221-8233); 1EZQ, 1FOR and 1FOS(Maignan, S., et al., J Med Chem., 2000, 43, 3226-3232); 1FJS (Adler,M., et al., Biochemistry, 2000, 39, 12534-12542); 1KSN (Guertin, K. R.,et al., Bioorg Med Chem Lett., 2002, 12, 1671-1674); 1NFU, 1NFW, 1NFXand 1NFY (Maignan, S., et al., J Med Chem., 2003, 46, 685-690); 2XBV,2XBW, 2XBX, 2XBY, 2XCO, 2XC4 and 2XC5 (Anselm, L., et al., Bioorg MedChem Lett., 2010, 20, 5313); 4A7I (Nazare, M., et al., Angew Chem Int EdEngl., 2012, 51, 905); 4BTI, 4BTT and 4BTU (Meneyrol, L., et al., J MedChem., 2013, 56, 9441); 3FFG, 3KQB, 3KQC, 3KQD and 3KQE (Quan, M. L., etal., Bioorg Med Chem Lett., 2010, 20, 1373-1377); 2P93, 2P94 and 2P95(Qiao, J. X., et al., Bioorg Med Chem Lett., 2007, 17, 4419-4427); 1V3X(Haginoya, N., et al., J Med Chem., 2004, 47, 5167-5182); 2P16 (Pinto,D. J. P., et al., J Med Chem., 2007, 50, 5339-5356); 2RAO (Lee, Y. K.,et al., J Med Chem., 2008, 51, 282-297); 3SW2 (Shi, Y., et al., BioorgMed Chem Lett., 2011, 21, 7516-7521); 2VH6 (Young, R. J., et al., BioorgMed Chem Lett., 2008, 18, 23); 2WYG and 2WYJ (Kleanthous, S., et al.,Bioorg Med Chem Lett., 2010, 20, 618); 2Y7X (Watson, N. S., et al.,Bioorg Med Chem Lett., 2011, 21, 1588); 2Y7Z, 2Y80, 2Y81 and 2Y82(Young, R. J., et al., Bioorg Med Chem Lett., 2011, 21, 1582); 3KL6(Fujimoto, T., et al., J Med Chem., 2010, 53, 3517-3531); 3LIW (Meuller,M. M., et al., Biol.Chem., 2003, 383, 1185); 5KOH (Schweinitz, A., etal., Med Chem., 2006, 2, 349-361); 1XKA and 1XKB (Kamata, K., et al.,Proc Natl Acad Sci USA, 1998, 95, 6630-6635); 2EI6 and 2EI7 (Nagata, T.,et al., Bioorg Med Chem Lett., 2007, 17, 4683-4688); 2P3T (Ye, B., etal., J Med Chem., 2007, 50, 2967-2980); 1MQ5 and 1MQ6 (Adler, M., etal., Biochemistry, 2002, 41, 15514-15523); 3K9X and 3HPT (Shi, Y., etal., Bioorg Med Chem Lett., 2009, 19, 6882-6889); 3CEN (Corte, J. R., etal., Bioorg Med Chem Lett., 2008, 18, 2845-2849); 2W3I and 2W3K (VanHuis, C. A., et al., Bioorg Med Chem., 2009, 17, 2501); 2H9E (Murakami,M. T., et al., J Mol Biol., 2007, 366, 602-610); 1WU1 and 2D1J(Komoriya, S., et al., Bioorg Med Chem., 2005, 13, 3927-3954); 2G00(Pinto, D. J. P., et al., Bioorg Med Chem Lett., 2006, 16, 5584-5589);3M36 and 3M37 (Pruitt, J. R. et al., J Med Chem., 2003, 46, 5298-5315);3CS7 (Qiao, J. X., et al., Bioorg Med Chem Lett., 2008, 18, 4118-4123);1Z6E (Quan, M. L., et al., J Med Chem., 2005, 48, 1729-1744); 2FZZ(Pinto, D. J. P., et al., Bioorg Med Chem Lett., 2006, 16, 4141-4147);and 3ENS (Shi, Y., et al., J Med Chem., 2008, 51, 7541-7551).

Representative Factor Xa Targeting Ligands are provided in FIG. 1 .Additional Factor Xa Targeting Ligands can be found in, for example,Bioorg Med Chem Lett 20: 5313-9 (2010), Bioorg Med Chem Lett 13: 679-83(2003), J Med Chem 44: 566-78 (2001), J Med Chem 50: 2967-80 (2007), JMed Chem 38: 1511-22 (1995), Bioorg Med Chem Lett 18: 2845-9 (2008), JMed Chem 53: 6243-74 (2010), Bioorg Med Chem Lett 18: 2845-9 (2008),Bioorg Med Chem 16: 1562-95 (2008), each of which is incorporated hereinby reference.

Factor XI

In some embodiments, the Target Protein is human Factor XIUniProtKB—P03951 (FA11_HUMAN). Factor XI triggers the middle phase ofthe intrinsic pathway of blood coagulation by activating factor IX.

Factor XI has been implicated in the development of deep vein thrombosisand acute pulmonary embolism, and the risk of stroke and embolism inpeople with nonvalvular atrial fibrillation.

The Protein Data Bank website provides the crystal structure of FactorXI bound to various compounds searchable by 1ZSL, 1ZTJ, 1ZTK, and 1ZTL(Nagafuji, P., et al.,); 1ZOM (Lin, J., et al., J Med Chem., 2006, 49,7781-7791); 5EOK and 5EOD (Wong, S. S., et al., Blood, 2016, 127,2915-2923); 1ZHM, 1ZHP and 1ZHR (Jin, L., et al., Acta Crystallogr DBiol Crystallogr., 2005, 61, 1418-1425); 1ZMJ, 1ZLR, 1ZML and 1ZMN(Lazarova, T.I., Bioorg Med Chem Lett., 2006, 16, 5022-5027); 1ZRK, 1ZSJand 1ZSK (Guo, Z., et al); 4CRA, 4CRB, 4CRC, 4CRD, 4CRE, 4CRF and 4CRG(Fjellstrom, 0, et al., PLoS One, 2015, 10, 13705); 3SOR and 3SOS(Fradera, X., et al., Acta Crystallogr Sect F Struct Biol Cryst Commun.,2012, 68, 404-408); 1ZPB, 1ZPC, 2FDA (Deng, H., et. al., Bioorg Med ChemLett., 2006, 16, 3049-3054); 5WB6 (Wang, C., et al., Bioorg Med ChemLett.,2017, 27, 4056-4060); 4NA7 and 4NA8 (Quan, M. L., et al., J MedChem., 2014, 57, 955-969); 4WXI (Corte, J. R., et al., Bioorg Med ChemLett., 2015, 25, 925-930); 5QTV, 5QTW, 5QTX and 5QTY (Fang, T., et al.,Bioorg Med Chem Lett., 2020, 126949-126949); 6COS (Hu, Z., et al.,Bioorg Med Chem Lett., 28, 987-992); 5QQP and 5QQO (Clark, C. G., etal., Bioorg Med Chem Lett., 2019, 29, 126604-126604); 5QOD, 5QOE, 5QOF,5QOG, and 5QOH (Corte, J. R., et al., Bioorg Med Chem Lett., 2017, 27,3833-3839); 5QCK, 5QCL, 5QCM, and 5QCN (Pinto, D. J. P., et al., J MedChem., 2017, 60, 9703-9723); 5TKS and 5TKU (Corte, J. R., et al., J MedChem., 2017, 60, 1060-1075); 1XXD and 1XX9 (Jin, L., et al., J BiolChem., 2005, 280, 4704-4712); 5QTT and 5QTU (Corte, J. R., et al., J MedChem., 2019, 63, 784-803); 4TY6, 4TY7 (Hangeland, J. J., et al., J MedChem., 2014, 57, 9915-9932); 4X6M, 4X6N, 4X60, and 4X6P (Pinto, D. J.P., et al., Bioorg Med Chem Lett., 2015, 25, 1635-1642); and 5EXM(Corte, J. R., et al., Bioorg Med Chem., 2016, 24, 2257-2272).Additionally, A1-Horani et al., provides insight into a review of patentliterature regarding Factor Xia inhibitors (A1-Horani et al., ExpertOpin Ther Pat. 2016; 26(3), 323-345).

Representative Factor XI Targeting Ligands are provided in FIG. 1 .Additional Factor XI Targeting Ligands can be found in, for example,U.S. Pat. No. 9,783,530, U.S. patent Ser. No. 10/143,681, U.S. patentSer. No. 10/214,512, ACS Med Chem Lett 6: 590-5 (2015), J Med Chem 60:9703-9723 (2017), J Med Chem 60: 9703-9723 (2017), U.S. Pat. No.9,453,018 (2016), J Med Chem 60: 1060-1075 (2017), J Med Chem 57: 955-69(2014), each of which is incorporated herein by reference.

Factor XII

In some embodiments, the Target Protein is human Factor XII(UniProtKB—P00748 (FA12_HUMAN)). Factor XII is a serum glycoprotein thatparticipates in the initiation of blood coagulation, fibrinolysis, andthe generation of bradykinin and angiotensin. Prekallikrein is cleavedby factor XII to form kallikrein, which then cleaves factor XII first toalpha-factor XIIa and then trypsin cleaves it to beta-factor XIIa.Alpha-factor XIIa activates factor XI to factor XIa.

Factor XII has been implicated in the development of deep veinthrombosis and acute pulmonary embolism, and the risk of stroke andembolism in people with nonvalvular atrial fibrillation.

The Protein Data Bank website provides the crystal structure of factorXII bound to various compounds searchable by 4XDE and 4XE4 (Pathak, M.,et al., J Thromb Haemost., 2015, 13(4), 580-591); 6GT6 and 6QF7 (Pathak,M., et al., Acta Crystallogr D Struct Biol., 2019, 75, 578-591); and6B74 and 6B77 (Dementiev, A. A., et al., Blood Adv., 2018, 2, 549-558).Additionally, Pathak et al., provides insight into the crystal structureof factor XII (Pathak, M., et al., J Thromb Haemost., 2015, 13(4),580-591).

Representative Factor XII Targeting Ligands are provided in FIG. 1 .Additional Factor XII Targeting Ligands can be found in, for example, JMed Chem 60: 1151-1158 (2017), J Med Chem 48: 2906-15 (2005), J Med Chem50: 5727-34 (2007), J Med Chem 50: 1876-85 (2007), Chembiochem 18:387-395 (2017), each of which is incorporated herein by reference.

Factor XIII

In some embodiments, the Target Protein is human Factor XIIIUniProtKB—P00488 (F13A_HUMAN)). Factor XIII is activated by thrombin andcalcium ion to a transglutaminase that catalyzes the formation ofgamma-glutamyl-epsilon-lysine cross-links between fibrin chains, thusstabilizing the fibrin clot. Also cross-link alpha-2-plasmin inhibitor,or fibronectin, to the alpha chains of fibrin.

Factor XIII has been implicated in the development of deep veinthrombosis and acute pulmonary embolism, and the risk of stroke andembolism in people with nonvalvular atrial fibrillation.

The Protein Data Bank website provides the crystal structure of factorXIII searchable by 1FIE (Yee, V. C., et al., Thromb Res., 1995, 78,389-397); and 1F13 (Weiss, M. S., et al., FEBS Lett., 1998, 423,291-296); as well as the crystal structure of factor XIII bound tovarious compounds searchable by 1DE7 (Sadasivan, C., et al., J BiolChem., 2000, 275, 36942-36948); and 5MHL, 5MHM, 5MHN, and 5MHO (Stieler,M., et al.,). Additionally, Gupta et al., provides insight into themechanism of coagulation factor XIII activation and regulation from astructure/functional perspective (Gupta, S., et al., Sci Rep., 2016; 6,30105); and Komaromi et al., provides insight into the novel structuraland functional aspect of factor XIII (Komaromi, Z., et al., J ThrombHaemost 2011, 9, 9-20).

Representative Factor XIII Targeting Ligands are provided in FIG. 1 .Additional Factor XIII Targeting Ligands can be found in, for example,Eur J Med Chem 98: 49-53 (2015), J Med Chem 55: 1021-46 (2012), J MedChem 48: 2266-9 (2005), each of which is incorporated herein byreference.

Prothrombin

In some embodiments, the Target Protein is human Prothrombin(UniProtKB—P00734 (THRB_HUMAN)). Thrombin, which cleaves bonds after Argand Lys, converts fibrinogen to fibrin and activates factors V, VII,VIII, XIII, and, in complex with thrombomodulin, protein C.

Functions in blood homeostasis, inflammation and wound healing. Thrombinis involved in blood clot formation and arterial and venous thrombosis,and thromboembolism associated with atrial fibrillation.

The Protein Data Bank website provides the crystal structure ofprothrombin searchable by 3NXP (Chen, Z. et al., Proc Natl Acad Sci USA,2010, 107, 19278-19283); as well as the crystal structure of prothrombinbound to various compounds searchable by 2HIPP and 2IPQ (Arni, R. K., etal., Biochemistry, 1993, 32, 4727-4737); 6BJR, 6C2W (Chinnaraj, M., etal., Sci Rep., 2018, 8, 2945-2945); 5EDK, 5EDM (Pozzi, N., et al., JBiol Chem., 2016, 291, 6071-6082); 3K65 (Adams, T. E., et al.,Biochimie, 2016, 122, 235-242); and 6BJR and 6C2W (Chinnaraj, M. et al.,Sci Rep., 2018, 8, 2945-2945). Additionally, Pozzi et al., providesinsight into the mechanism and conformational flexibility for thecrystal structure of prothrombin (Pozzi, N. et al., J Biol Chem., 2013,288(31), 22734-22744); and Zhiwei et al., provides insight into thecrystal structure of prothrombin-1 (Zhiwei, C. et al., PNAS, 2010,107(45), 19278-19283).

Prothrombin is converted to thrombin, as such the Protein Data Bankwebsite provides the crystal structure of thrombin bound to compoundssearchable by 1XMN (Carter, W. J. et al., J.Biol.Chem., 2005, 280,2745-2749); 4CH₂ and 4CH8 (Lechtenberg, B. C. et al., J Mol Biol., 2014,426, 881); 3P01 (Karle, M. et al., Bioorg Med Chem Lett., 2012, 22,4839-4843); 3DA9 (Nilsson, M. et al., J Med Chem., 2009, 52, 2708-2715);2H9T and 3BF6 (Lima, L. M. T. R. et al., Biochim Biophys Acta., 2009,1794, 873-881); 3BEF and 3BEI (Gandhi, P. S. et al., Proc Natl Acad SciUSA, 2008, 105, 1832-1837); 3BV9 (Nieman, M. T. et al., J ThrombHaemost., 2008, 6, 837-845); 2HWL (Pineda, A. O. et al., Biophys Chem.,2007, 125, 556-559); 2AFQ (Johnson, D. J. D. et al., Biochem J., 2005,392, 21-28); 1SHH (Pineda, A. O. et al., J Biol Chem., 2004, 279,31842-31853); 1JWT (Levesque, S. et al., Bioorg Med Chem Lett., 2001,11, 3161-3164); 1G37 (Bachand, B. et al., Bioorg Med Chem Lett., 2001,11, 287-290); 1EOJ and 1EOL (Slon-Usakiewicz, J. J. et al.,Biochemistry, 2000, 39, 2384-2391); 1AWH (Weir, M. P. et al.,Biochemistry, 1998, 37, 6645-6657); 1DIT (Krishnan, R. et al., ProteinSci., 1996, 5, 422-433); 1HAO and 1HAP (Padmanabhan, K. et al., ActaCrystallogr D Biol Crystallogr., 1996, 52, 272-282); and 1HBT (Rehse, P.H. et al., Biochemistry, 1995, 34, 11537-11544).

Representative prothrombin Targeting Ligands are provided in FIG. 1 .Additional prothrombin Targeting Ligands can be found in, for example, JMed Chem 46: 3612-22 (2003), Bioorg Med Chem Lett 12: 1017-22 (2002), JMed Chem 40: 830-2 (1997), Bioorg Med Chem Lett 15: 2771-5 (2005), J MedChem 42: 3109-15 (1999), J Med Chem 47: 2995-3008 (2004), Bioorg MedChem 16: 1562-95 (2008), J Med Chem 42: 3109-15 (1999), each of which isincorporated herein by reference.

Coagulation Factor VII

In some embodiments, the Target Protein is human coagulation Factor VII(UniProtKB-P08709 (FA7_HUMAN)). Factor VII initiates the extrinsicpathway of blood coagulation. It is a serine protease that circulates inthe blood in a zymogen form. Factor VII is converted to Factor VIIa byFactor Xa, Factor XIIa, Factor IXa, or thrombin by minor proteolysis. Inthe presence of tissue factor and calcium ions, Factor VIIa thenconverts Factor X to Factor Xa by limited proteolysis. Factor VIIa willalso convert Factor IX to Factor IXa in the presence of tissue factorand calcium.

Factor VII is involved in blood clot formation and arterial and venousthrombosis, and thromboembolism associated with atrial fibrillation.

The Protein Data Bank website provides the crystal structure of factorVII bound to various compounds searchable by 2F9B (Rai, R., et al.,Bioorg Med Chem Lett., 2006, 16, 2270-2273); 5U6J (Wurtz, N. R., et al.,Bioorg Med Chem Lett., 2017, 27, 2650-2654); 5L2Y, 5L2Z, and 5L30(Ladziata, U., et al., Bioorg Med Chem Lett., 2016, 26, 5051-5057); 5146(Glunz, P. W., et al., J Med Chem., 2016, 59, 4007-4018); 4YLQ, 4Z6A,and 4ZMA (Sorensen, A. B., et al., J Biol Chem., 2016, 291, 4671-4683);4YT6 and 4YT7 (Glunz, P. W., et al., Bioorg Med Chem Lett, 2015, 25,2169-2173); 4NA9 (Quan, M. L., et al., J Med Chem., 2014, 57, 955-969);4NG9 (hang, X., et al., ACS Med Chem Lett., 2014, 5, 188-192); 4JZD,4JZE and 4JZF (Bolton, S. A., et al., Bioorg Med Chem Lett., 2013, 23,5239-5243); 4JYU and 4JYV (Glunz, P. W., et al., Bioorg Med Chem Lett.,2013, 23, 5244-5248); 4ISH (Priestley, E. S., et al., Bioorg Med ChemLett., 2013, 23, 2432-2435); 4ISI (Zhang, X., et al., Bioorg Med ChemLett., 2013, 23, 1604-1607); 2ZZU (Shiraishi, T., et al., Chem PharmBull (Tokyo), 2010, 58, 38-44); 1WV7 and 1WUN (Kadono, S., et al.,Biochem Biophys Res Commun., 2005, 327, 589-596); 2ZWL, 2ZPO, (Kadono,S., et al.); 2EC9 (Krishan, R., et al., Acta Crystallogr D BiolCrystallogr., 2007, 63, 689-697); 2PUQ (Larsen, K. S., et al., BiochemJ., 2007, 405, 429-438); 2FLR (Riggs, J. R., et al., Bioorg Med ChemLett., 2006, 16, 3197-3200); 2C4F (Kohrt, J. T., et al., Bioorg Med ChemLett., 2006, 16, 1060); 2AEI (Kohrt, J. T. et al., Bioorg Med ChemLett., 2005, 15, 4752-4756); 1WTG (Kadono, S., et al., Biochem BiophysRes Commun., 2005, 326, 859-865); 1WSS (Kadono, S., et al., ActaCrystallogr Sect F Struct Biol Cryst Commun., 2005, 61, 169-173); 1W7Xand 1W8B (Zbinden, K. G., et al., Bioorg Med Chem Lett., 2005, 15,5344); 1WQV (Kadono, S., et al., Biochem Biophys Res Commun., 2004, 324,1227-1233); 1Z6J (Schweitzer, B. A., et al., Bioorg Med Chem Lett.,2005, 15, 3006-3011); 1YGC (Olivero, A. G., et al., J Biol Chem., 2005,280, 9160-9169); 6R2W (Sorensen, A. B., et al., J Biol Chem., 2019, 295,517-528); 5PA8, 5PA9, 5PAA, 5PAB, 5PAC, 5PAE, 5PAF, 5PAG, 5PAI, 5PAJ,5PAK, 5PAM, 5PAN, 5PAO, 5PAQ, 5PAR, 5PAS, 5PAT, 5PAU, 5PABV, 5PAW, 5PAX,5PAY, 5PBO, 5PB1, 5PB2, 5PB3, 5PB4, 5PB5, and 5PB6 (Mayweg, A. V., etal.,); and 5LOS (Li, Z., et al., Nat Commun., 2017, 8, 185-185).Additionally, Kemball-Cook, et al., provides insight into the crystalstructure of active site-inhibited factor VIIa (Kemball-Cook, G., etal., J Struct Biol., 1999, 127(3), 213-23).

Representative Factor VII Targeting Ligands are provided in FIG. 1 .Additional Factor VII Targeting Ligands can be found in, for example,U.S. Pat. No. 9,174,974, Bioorg Med Chem Lett 26: 5051-5057 (2016),Bioorg Med Chem Lett 11: 2253-6 (2001), Bioorg Med Chem Lett 15: 3006-11(2005), Bioorg Med Chem Lett 12: 2883-6 (2002), each of which isincorporated herein by reference.

Coagulation Factor IX

In some embodiments, the Target Protein is human coagulation Factor IX(UniProtKB-P00740 (FA9_HUMAN)). Factor IX Factor IX is a vitaminK-dependent plasma protein that participates in the intrinsic pathway ofblood coagulation by converting factor X to its active form in thepresence of Ca2+ ions, phospholipids, and factor VIIIa.

Factor IX is involved in blood clot formation and arterial and venousthrombosis, and thromboembolism associated with atrial fibrillation.

The Protein Data Bank website provides the crystal structure of factorIX bound to various compounds searchable by 6MV4 (Vadivel, K., et al., JThromb Haemost., 2019, 17, 574-584); 4ZAE (Zhang, T., et al., Bioorg MedChem Lett., 2015, 25, 4945-4949); 4YZU and 4ZOK (Parker, D. L., et al.,Bioorg Med Chem Lett., 2015, 25, 2321-2325); 5TNO and 5TNT (Sakurada,I., et al., Bioorg Med Chem Lett., 2017, 27, 2622-2628); 5JB8, 5JB9,5JBA, 5JBB and 5JBC (Kristensen, L. H., et al., Biochem J., 2016, 473,2395-2411); 3LC3 (Wang, S., et al., J Med Chem., 2010, 53, 1465-1472);3LC5 (Wang, S., et al., J Med Chem., 2010, 53, 1473-1482); 3KCG(Johnson, D. J. D., et al., Proc Natl Acad Sci USA, 2010, 107, 645-650);1NLO (Huang, M., et al., J Biol Chem., 2004, 279, 14338-14346); 1RFN(Hopfner, K. P., et al., Structure, 1999, 7, 989-996); and 6RFK(Sendall, T. J., et al.,).

Representative Factor IX Targeting Ligands are provided in FIG. 1 .Additional Factor IX Targeting Ligands can be found in, for example,U.S. Pat. No. 9,409,908, Bioorg Med Chem Lett 25: 5437-43 (2015), U.S.patent Ser. No. 10/189,819, each of which is incorporated herein byreference.

Fibroblast Growth Factor 1 (FGF1)

In some embodiments, the Target Protein is human fibroblast growthfactor 1 (FGF1) (UniProtKB—P05230 (FGF1_HUMAN)). FGF1 plays an importantrole in the regulation of cell survival, cell division, angiogenesis,cell differentiation and cell migration. FGF1 acts as a ligand for FGFR1and integrins, and binds to FGFR1 in the presence of heparin leading toFGFR1 dimerization and activation via sequential autophosphorylation ontyrosine residues which act as docking sites for interacting proteins,leading to the activation of several signaling cascades. FGF1 inducesthe phosphorylation and activation of FGFR1, FRS2, MAPK3/ERK1,MAPK1/ERK2 and AKT1. FGF1 can induce angiogenesis. FGF1 has beenimplicated in oncogenesis, cancer cell proliferation, resistance toanticancer therapies, and neoangiogenesis.

The Protein Data Bank website provides the crystal structure of FGF1searchable by 2AFG (Blaber, M., et al., Biochemistry, 1996, 35,2086-2094); and 1BAR (Zhu, X. et al., Science, 1991, 251, 90-93); aswell as the crystal structure of FGF1 bound to various compoundssearchable by 1AFC (Zhu, X., et al., Structure, 1993, 1, 27-34); 1AXMand 2AXM (DiGabriele, A. D., et al., Nature, 1998, 393, 812-817); IEVT(Plotnikov, A. N., et al., Cell, 2000, 101, 413-424); 1E0O (Pellegrini,L., et al., Nature, 2000, 407, 1029); and 2ERM (Canales, A., et al.,FEBS J, 2006, 273, 4716-4727).

Representative FGF1 Targeting Ligands are provided in FIG. 1 .Additional FGF1 Targeting Ligands can be found in, for example, BioorgMed Chem Lett 18: 344-9 (2008), Chembiochem 6: 1882-90 (2005), J MedChem 55: 3804-13 (2012), J Med Chem 47: 1683-93 (2004), J Med Chem 53:1686-99 (2010,)each of which is incorporated herein by reference.

Fibroblast Growth Factor 2 (FGF2)

In some embodiments, the Target Protein is human fibroblast growthfactor 2 (FGF2) (UniProtKB—P09038 (FGF2_HUMAN)). FGF2 acts as a ligandfor FGFR1, FGFR2, FGFR3 and FGFR4. FGF2 also acts as an integrin ligandwhich is required for FGF2 signaling, and plays an important role in theregulation of cell survival, cell division, cell differentiation andcell migration. FGF2 also induces angiogenesis. FGF2 has been implicatedin oncogenesis, cancer cell proliferation, resistance to anticancertherapies, and neoangiogenesis.

The Protein Data Bank website provides the crystal structure of FGF2bound to various compounds searchable by 40EE, 40EF, and 40EG (Li, Y.C., et al., ACS Chem Biol., 2014, 9, 1712-1717); 1EV2 (Plotnikov, A. N.,et al., Cell, 2000, 101, 413-424); and 5X10 (Tsao, Y. H.).

Representative FGF2 Targeting Ligands are provided in FIG. 1 .Additional FGF2 Targeting Ligands can be found in, for example, U.S.Pat. No. 8,933,099, Bioorg Med Chem Lett 12: 3287-90 (2002), Chem BiolDrug Des 86: 1323-9 (2015), Bioorg Med Chem Lett 25: 1552-5 (2015), eachof which is incorporated herein by reference.

Fibronectin-1

In some embodiments, the Target Protein is human fibronectin 1 (FN1)(UniProtKB-P02751 (FINC_HUMAN)). Fibronectin (FN) polymerization isnecessary for collagen matrix deposition and is a key contributor toincreased abundance of cardiac myofibroblasts (MFs) after cardiacinjury. Interfering with FN polymerization may attenuate MF and fibrosisand improve cardiac function after ischemia/reperfusion (I/R) injury.

The Protein Data Bank website provides the crystal structure offibronectin-1 bound to various compounds searchable by 3M7P (Graille,M., et al., Structure, 2010, 18, 710-718); 3MQL (Erat, M. C., et al., JBiol Chem., 2010, 285, 33764-33770); and 3EJH (Erat, M. C., et al., ProcNatl Acad Sci USA, 2009, 106, 4195-4200).

Representative FN Targeting Ligands are provided in FIG. 1 . AdditionalFN Targeting Ligands can be found in, for example, Bioorg Med Chem Lett18: 2499-504 (2008), which is incorporated herein by reference.

Kallikrein-1 (KLK1)

In some embodiments, the Target Protein is human kallikrein-1(UniProtKB—P06870 (KLK1_HUMAN)). Glandular kallikreins cleave Met-Lysand Arg-Ser bonds in kininogen to release Lys-bradykinin. Kallikrein hasbeen implicated in adverse reactions in hereditary angioedema (HAE).

The Protein Data Bank website provides the crystal structure of KLK1searchable by 1SPJ (Laxmikanthan, G., et al., Proteins, 2005, 58,802-814); as well as the crystal structure of KLK1 bound to variouscompounds searchable by 5F8Z, 5F8T, 5F8X, (Xu, M., et al.,); and 6A80(Xu, M., et al., FEBS Lett., 2018, 592, 2658-2667). Additionally, Katzet al., provides insight into the crystal structure of kallikrein (Katz,B. A., et al., Protein Sci., 1998, 7(4), 875-85).

Representative kallikrein Targeting Ligands are provided in FIG. 1 .Additional kallikrein Targeting Ligands can be found in, for example,U.S. Pat. No. 9,783,530, J Med Chem 38: 2521-3 (1995), U.S. Pat. No.9,234,000, U.S. patent Ser. No. 10/221,161, U.S. Pat. Nos. 9,687,479,9,670,157, 9,834,513, J Med Chem 38: 1511-22 (1995), U.S. patent Ser.No. 10/214,512, each of which is incorporated herein by reference.

Plasma Kallikrein

In some embodiments, the Target Protein is human plasma kallikrein(UniProtKB—P03952 (KLKB1_HUMAN)). Plasma kallikrein cleaves Lys-Arg andArg-Ser bonds. It activates, in a reciprocal reaction, factor XII afterits binding to a negatively charged surface. It also releases bradykininfrom HMW kininogen and may also play a role in the renin-angiotensinsystem by converting prorenin into renin. Plasma kallikrein has beenimplicated in retinal dysfunction, the development of diabetic macularedema and hereditary angioedema (HAE).

The Protein Data Bank website provides the crystal structure of plasmakallikrein bound to various compounds searchable by 5TJX (Li, Z., etal., ACS Med Chem Lett., 2017, 8, 185-190); 601G and 601S (Patridge, J.R., et al., J Struct Biol., 2019, 206, 170-182); 40GX and 40GY(Kenniston, J. A., et al., J Biol Chem., 2014, 289, 23596-23608); and5F8T, 5F8X, and 5F8Z (Xu, M., et al.,).

Representative plasma kallikrein Targeting Ligands are provided in FIG.1 . Additional plasma kallikrein Targeting Ligands can be found in, forexample, J Med Chem 61: 2823-2836 (2018), J Med Chem 55: 1171-80 (2012),U.S. Pat. Nos. 8,598,206, 9,738,655, Bioorg Med Chem Lett 16: 2034-6(2006), U.S. Pat. No. 9,409,908, U.S. patent Ser. No. 10/144,746, U.S.Pat. No. 9,290,485, each of which is incorporated herein by reference.

Lipoprotein Lipase

In some embodiments, the Target Protein is human lipoprotein lipase(UniProtKB—P06858 (LIPL_HUMAN)). Lipoprotein lipase is a key enzyme intriglyceride metabolism. It catalyzes the hydrolysis of triglyceridesfrom circulating chylomicrons and very low density lipoproteins (VLDL),and thereby plays an important role in lipid clearance from the bloodstream, lipid utilization and storage. Lipoprotein lipase mediatesmargination of triglyceride-rich lipoprotein particles in capillaries.Lipoprotein lipase has been implicated in the development ofcardiovascular disease and obesity.

The Protein Data Bank website provides the crystal structure oflipoprotein lipase bound to various compounds searchable by 6E7K(Birrane, G., et al., Proc Natl Acad Sci USA, 2018 116 1723-1732).

Representative lipoprotein lipase Targeting Ligands are provided in FIG.1 . Additional lipoprotein lipase Targeting Ligands can be found in, forexample, J Med Chem 47: 400-10 (2004), which is incorporated herein byreference.

Matrix Metallopeptidase 1 (MMP-1)

In some embodiments, the Target Protein is human matrix metallopeptidase1 (MMP-1) (UniProtKB—P03956 (MMP1_HUMAN)). MMP-1 cleaves collagens oftypes I, II, and III at one site in the helical domain. It also cleavescollagens of types VII and X. MMP-1 has been implicated incardiovascular disease.

The Protein Data Bank website provides the crystal structure of MMP-1searchable by 3SHI (Bertini, I., et al., FEBS Lett., 2012, 586,557-567); as well as the crystal structure of MMP-1 bound to variouscompounds searchable by 4AUO (Manka, S. W., et al., Proc Natl Acad SciUSA, 2012, 109, 12461); 3MA2 (Grossman, M., et al., Biochemistry, 2010,49, 6184-6192); and 2JOT (Iyer, S., et al., J.Biol.Chem., 2007, 282,364). Additionally, Iyer et al., provides insight into the crystalstructure of an active form of MMP-1 (Iyer, S., et al., J Mol Biol.,2006, 362(1), 78-88); and Lovejoy et al., provides insight into thecrystal structure of MMP1 and the selectivity of collagenase inhibitors(Lovejoy, B., et al., Nat Struct Mol Biol., 1999, 6, 217-221).

Representative MMP-1 Targeting Ligands are provided in FIG. 1 .Additional MMP-1 Targeting Ligands can be found in, for example, BioorgMed Chem Lett 5: 1415-1420 (1995), Bioorg Med Chem Lett 16: 2632-6(2006), Bioorg Med Chem Lett 8: 837-42 (1999), Eur J Med Chem 60: 89-100(2013), J Med Chem 54: 4350-64 (2011), Bioorg Med Chem Lett 8: 3251-6(1999), J Med Chem 42: 4547-62 (1999), J Med Chem 61: 2166-2210 (2018),J Med Chem 41: 1209-17 (1998), which is incorporated herein byreference.

Macrophage Migration Inhibitory Factor (MIF)

In some embodiments, the Target Protein is human macrophage migrationinhibitory factor (MIF) (UniProtKB—P14174 (MIF_HUMAN)). MIF is apro-inflammatory cytokine involved in the innate immune response tobacterial pathogens. The expression of MIF at sites of inflammationsuggests a role as mediator in regulating the function of macrophages inhost defense. It counteracts the anti-inflammatory activity ofglucocorticoids.

MIF has been implicated in tumor progression; systemic inflammation;atherosclerosis; rheumatoid arthritis; and systemic lupus erythematosus,among others.

The Protein Data Bank website provides the crystal structure of MIFsearchable by 1MIF (Sun, H-W. et al., Proc Natl Acad Sci USA, 1996, 93,5191-5196); as well as the crystal structure of MIF bound to variouscompounds searchable by 6PEG (Cirillo, P. F. et al.,); 5XEJ (Fukushima,K); 6FVE and 6FVH (Sokolov, A. V., et al., Biochemistry (Mosc), 2018,83, 701-707); 6CB5, 6CBF, 6CBG, and 6CBH (Trivedi-Parmar, V., et al.,ChemMedChem., 2018, 13, 1092-1097); 6B1C, 6B1K, 6B2C, (Dawson, T. K., etal., ACS Med Chem Lett., 2017, 8, 1287-1291); 4Z15, 4Z1T and 4Z1U(Singh, A. K., et al, J Cell Mol Med., 2017, 21, 142-153); 5HVS and 5HVT(Cisneros, J. A., et al., J Am Chem Soc., 2016, 138, 8630-8638); 4PKK(Pantouris, G., et al.,); 5J7P and 5J7Q (Cisneros, J. A., et al., BioorgMed Chem Lett., 2016, 26, 2764-2767); 5B40 (Kimura, H., et al., ChemBiol., 2010, 17, 1282-1294); 4PLU, 4TRF, 4POH, and 4P01 (Pantouris, G.,et al., Chem Biol., 2015, 22, 1197-1205); 4WR8 and 4WRB (Dziedzic, P.,et al., J Am Chem Soc., 2015, 137 2996-3003); 4K9G (Ioannou, K., etal.,Int J Oncol., 2014, 45, 1457-1468); 40SF, 3WNR, 3WNS and 3WNT (Spencer,E. S., et al., Eur J Med Chem., 2015, 93, 501-510); 40YQ (Spencer, E. S.et al.,); 3SMB and 3SMC (Crichlow, G. V. et al., Biochemistry, 2012, 51,7506-7514); 3U18 (Bai, F., et al., J Biol Chem., 2012, 287,30653-30663); 4F2K (Tyndall, J. D. A., et al., Acta Crystallogr Sect FStruct Biol Cryst Commun., 2012, 68, 999-1002); 3IJG and 3IJJ (Cho, Y.,et al., Proc Natl Acad Sci USA, 2010, 107, 11313-11318); 3L5P, 3L5R,3L5S, 3L5T, 3L5U, and 3L5V (McLean, L. R. et al., Bioorg Med Chem Lett.,2010, 20, 1821-1824); 3JSF, 3JSG and 3JTU (McLean, L. R., et al., BioorgMed Chem Lett., 2009, 19, 6717); 3HOF (Crawley, L., et al.); 3CE4 and3DJI (Crichlow G.V., et al., Biochemistry, 2009, 48, 132-139); 3B9S(Winner, M. et al., Cancer Res., 2008, 68, 7253-7257); 200H, 20OW and200Z (Crichlow, G. V. et al., J Biol Chem., 2007, 282, 23089-23095);1GCZ and 1GDO (Orita, M. et al., J Med Chem., 2001, 44, 540-547); and1CA7, 1CGQ and 1P1G (Lubetsky, J. B. et al., Biochemistry, 1999, 38,7346-7354). Additionally, Sun et al., provides insight into the crystalstructure of MIF (Proc Natl Acad Sci USA, 1996, 28;93(11), 5191-6).

Representative MIF Targeting Ligands are provided in FIG. 1 . AdditionalMIF Targeting Ligands can be found in, for example, ACS Med Chem Lett 8:124-127 (2017), J Med Chem 44: 540-7 (2001), J Med Chem 52: 416-24(2009), J Med Chem 50: 1993-7 (2007), which is incorporated herein byreference.

Transforming Growth Factor-β2 (TGF-β2)

In some embodiments, the Target Protein is human transforming growthfactor-02 (TGF-02) (UniProtKB—P61812 (TGFB2_HUMAN)). TGF-β2 is amultifunctional protein that regulates various processes such asangiogenesis and heart development. Once activated following release ofLAP, TGF-beta-2 acts by binding to TGF-beta receptors (TGFBR1 andTGFBR2), which transduce signal. TGF-β2 expression in the tumormicroenvironment has been associated with a poor prognosis, and isimplicated in TGF-β2 mediated tumor suppression via T-cell exclusion.TGF-β2 expression has also been implicated in hematological malignanciesand fibrosis.

The Protein Data Bank website provides the crystal structure of TGF-β2searchable by 6I9J (Del Amo-Maestro L. et al., Sci Rep. 2019, 9,8660-8660); as well as the crystal structure of TGF-32 bound to variouscompounds searchable by 1M9Z (Boesen, C. C., et al. Structure, 2002, 10,913-919); 5QIN (Zhang, Y. et al., ACS Med Chem Lett., 2018, 9,1117-1122); 5E8V, 5E8Y, 5E91 and 5E92 (Tebben, A. J. et al., ActaCrystallogr D Struct Biol., 2016, 72, 658-674); 4P7U (Wangkanont, K. etal., Protein Expr Purif, 2015, 115, 19-25); 4XJJ (Wangkanont et al.);and 1KTZ (Hart, P. J., et al., Nat Struct Biol., 2002, 9, 203-208).

Representative TGF-β2 Targeting Ligands are provided in FIG. 1 .

Thrombospondin-1 (TSP-1)

In some embodiments, the Target Protein is human thrombospondin-1(TSP-1) (UniProtKB—P61812 (TGFB2_HUMAN)). TSP1 acts as an angiogenesisinhibitor by stimulating endothelial cell apoptosis, inhibitingendothelial cell migration and proliferation, and regulating vascularendothelial growth factor bioavailability and activity. TSP1 affectstumor immune response, tumor cell behaviors including adhesion,invasion, migration, apoptosis, and proliferation.

TSP-1 expression has been implicated in a number of diseases, includingin promoting certain cancers such as breast cancer, prostate cancer,melanoma, SCLC, osteosarcoma, cutaneous squamous cell carcinoma, oralsquamous cell carcinoma, papillary thyroid carcinoma, thyroid cancer,medulloblastoma, and fibrotic disorders such as diabetes, liverfibrosis, and in multiple myeloma.

The Protein Data Bank website provides the crystal structure of TSP-1searchable by 1LSL (Tan, K. et al., J Cell Biol., 2002, 159, 373-382);2ES3 (Tan, K., et al., J Biol Chem., 2008, 283, 3932-3941); 1Z78 and2ERF (Tan, K., et al., Structure, 2006, 14, 33-42); and 3R6B (Klenotic,P. A., et al., Protein Expr Purif., 2011, 80, 253-259); as well as thecrystal structure of TSP-1 bound to various compounds searchable by 20UHand 2OUJ (Tan, K., et al., J Biol Chem., 2008, 283, 3932-3941); and 1ZA4(Tan, K., et al., Structure, 2006, 14, 33-42).

Representative TSP-1 Targeting Ligands are provided in FIG. 1 .

CD40 Ligand (CD40L)

In some embodiments, the Target Protein is human CD40 ligand (CD40L)(UniProtKB-P29965 (CD40L_HUMAN)). CD40L is a cytokine that acts as aligand to CD40/TNFRSF5. It costimulates T-cell proliferation andcytokine production. Its cross-linking on T-cells generates acostimulatory signal which enhances the production of IL4 and IL10 inconjunction with the TCR/CD3 ligation and CD28 costimulation. CD40Linduces the activation of NF-kappa-B, as well as kinases MAPK8 and PAK2in T-cells. It also induces tyrosine phosphorylation of isoform 3 ofCD28. CD40L mediates B-cell proliferation in the absence of co-stimulusas well as IgE production in the presence of IL4, and is involved inimmunoglobulin class switching.

The Protein Data Bank website provides the crystal structure of CD40Lsearchable by 1ALY (Karpusas, M., et al., Structure, 1995, 3,1031-1039); as well as the crystal structure of CD40L bound to variouscompounds searchable by 3QD6 (An, H.J., et al., J Biol Chem., 2011, 286,11226-11235); and 6BRB (Karnell, J. L., et al., Sci Transl Med., 2019,11(489), 6584).

The expression of CD40L has been implicated in HIV-associatedneurocognitive disorders and cardiovascular complications.Representative CD40L Targeting Ligands are provided in FIG. 1 .

Urokinase-type Plasminogen Activator (UPA)

In some embodiments, the Target Protein is human urokinase-typeplasminogen activator (UPA) (UniProtKB—P00749 (UROK_HUMAN)).Urokinase-type plasminogen activator (uPA), is a serine protease presentin the blood and in the extracellular matrix of many tissues. Theprimary physiological substrate of this enzyme is plasminogen, which isan inactive form (zymogen) of the serine protease plasmin. Activation ofplasmin triggers a proteolytic cascade that, depending on thephysiological environment, participates in thrombolysis or extracellularmatrix degradation. This cascade had been involved in vascular diseasesand cancer progression. Elevated expression levels of urokinase andseveral other components of the plasminogen activation system are foundto be correlated with tumor malignancy.

The Protein Data Bank website provides the crystal structure of UPAbound to various compounds searchable by 5ZA7, 5ZAJ, 5ZA8, 5ZA9, 5ZAE,5ZAF, 5ZAG, 5ZAH, and 5ZC5 (Buckley, B. J. et al., J Med Chem., 2018,61, 8299-8320); 5LHP, 5LHQ, 5LHR, and 5LHS (Kromann-Hansen, T. et al.,Sci Rep., 2017, 7, 3385-3385); 2VNT (Fish, P. V. et al. J Med Chem.,2007, 50, 2341); 1OWD, 10WE, 1OWH, 1OWI, 1OWJ, and 1OWK (Wendt, M. D. etal., J Med Chem., 2004, 47, 303-324); 1SQA, iSQO, and 1SQT (Wendt, M.D., et al., Bioorg Med Chem Lett., 2004, 14, 3063-3068); 1U6Q (Bruncko,M. et al., Bioorg Med Chem Lett., 2005, 15, 93-98); 30X7, 30Y5 and 30Y6(Jiang, L. G. et al., J Mol Biol., 2011, 412, 235-250); 40S1, 40S2,40S4, 40S5, 40S6 and 40S7 (Chen, S. et al., Nat Chem., 2014, 6,1009-1016); 3IG6 (West, C. W. et al., Bioorg Med Chem Lett., 2009, 19,5712-5715); 4XOW and 4X1P (Jiang, L. et al., Int J Biochem Cell Biol.,2015, 62, 88-92); 4X1N, 4X1Q, 4X1R and 4X1S (Zhao, B. et al., PLoS One,2014, 9, e115872-e115872); 5WXO and 5WXP (Jiang, L. et al., BiochimBiophys Acta., 2018, 1862, 2017-2023); 4MNV, 4MNW, 4MNX, and 4MNY (Chen,S., et al., Angew Chem Int Ed Engl., 2014, 53, 1602-1606); 4GLY (Chen,S., et al., J Am Chem Soc., 2013, 135, 6562-6569); 4JK5 and 4JK5 (Chen,S., et al., Chembiochem., 2013, 14, 1316-1322); 3QN7 (Angelini, A. etal., ACS Chem Biol., 2012, 7, 817-821); 2NWN (Zhao, G. et al., J StructBiol., 2007, 160, 1-10); 6NMB (Wu, G. et al., Blood Adv., 2019, 3,729-733); 1WOZ, iW10, iW 11, 1W12, 1W13, and 1W14 (Zeslawska, E. et al.,J Mol Biol., 2003, 328, 109); 4DVA (Jiang, L et al., Biochem J., 2013,449, 161-166); 6A8G 6A8N (Wang, D. et al., J Med Chem., 2019, 62,2172-2183); 2VIN, 2VIO, 2VIP, 2VIQ, 2VIV, and 2VIW (Frederickson, M. etal., J Med Chem., 2008, 51, 183); 1EJN (Speri, S., et al., Proc NatlAcad Sci USA, 2000, 97, 5113-5118); 3PB1 (Lin, Z. et al., J Biol Chem.,2011, 286, 7027-7032); 3U73 (Xu, X. et al., J Mol Biol., 2012, 416,629-641); 1C5W, 1C5X, 1C5Y and ICSZ (Katz, B. A., et al., Chem Biol.,2000, 7, 299-312); 5XG4 (Xue, G. et al., Food Funct., 2017, 8,2437-2443); 5WXF (Jiang, L. et al., Biochim Biophys Acta., 2018, 1862,2017-2023); 5WXS, 4ZKS, 5WXQ, 5WXT, 5YC6, 5YC7, 5Z1C, (Jiang, L. etal.); 4H42 (Yu, H.Y. et al.,); 6AG3 and 6AG9 (Buckley, B. et al); 3KGP,3KHV, 3KID, 3M61, 3MHW, and 3MWI (Jiang, L. G. et al.,); 4ZKN, 4ZKO and4ZKR (Jiang, L. et al.); 208T, 208U, 208W (Zhao, G. et al.,); and 4FU7,4FU8, 4FU9, 4FUB, 4FUC, 4FUD, 4FUE, 4FUF, 4FUG, 4FUH, 4FUI, and 4FUJ(Kang, Y. N. et al.).

Representative UPA Targeting Ligands are provided in FIG. 1 . AdditionalUPA Targeting Ligands are provided in, for example, J Med Chem 38:1511-22 (1995), Bioorg Med Chem Lett 11: 2253-6 (2001), Bioorg Med ChemLett 14: 3063-8 (2004), J Med Chem 52: 3159-65 (2009), CSAR 1: (2012),Bioorg Med Chem 22: 3187-203 (2014), J Med Chem 50: 2341-51 (2007), JMol Biol 329: 93-120 (2003), Bioorg Med Chem Lett2:1399-1404 (1992), JMed Chem 35: 4297-305 (1992), J Med Chem 35: 4150-9 (1992), J Med Chem49: 5785-93 (2006), Bioorg Med Chem 23: 3696-704 (2015), Bioorg Med ChemLett 10: 983-7 (2000), J Med Chem 49: 5785-93 (2006), each of which isincorporated by reference herein.

Plasminogen Activator, Tissue Type (TPA)

In some embodiments, the Target Protein is human plasminogen activator,tissue type (TPA) (UniProtKB—P00750 (TPA_HUMAN)). TPA converts theabundant, but inactive, zymogen plasminogen to plasmin by hydrolyzing asingle Arg-Val bond in plasminogen. By controlling plasmin-mediatedproteolysis, it plays an important role in tissue remodeling anddegradation, in cell migration and many other physiopathological events.TPA plays a direct role in facilitating neuronal migration. PLA has beenshown activated in various cancers including oral malignancy.

The Protein Data Bank website provides the crystal structure of TPAsearchable by 1VR1 (Dekker, R. J. et al., J Mol Biol., 1999, 293,613-627); as well as the crystal structure of TPA bound to variouscompounds searchable by 1RTF (Lamba, D. et al., J Mol Biol., 1996, 258,117-135); 1ASH (Renatus, M. et al., J Biol Chem., 1997, 272,21713-21719); and 1BDA (Renatus, M. et al., EMBO J., 1997, 16,4797-4805).

Representative TPA Targeting Ligands are provided in FIG. 1 . AdditionalTPA Targeting Ligands are provided in, for example, Bioorg Med Chem Lett15: 4411-6 (2005), Bioorg Med Chem Lett 13: 2781-4 (2003), Bioorg MedChem Lett 6: 2913-2918 (1996), J Med Chem 44: 2753-71 (2001), J Med Chem41: 5445-56 (1999), Bioorg Med Chem Lett 12: 3183-6 (2002), U.S. patentSer. No. 10/118,930, J Biol Chem 285: 7892-902 (2010), each of which isincorporated by reference herein.

Plasminogen (PLG)

In some embodiments, the Target Protein is human plasminogen (PLG)(UniProtKB-P00747 (PLMN_HUMAN)). PLG dissolves the fibrin of blood clotsand acts as a proteolytic factor in a variety of other processesincluding embryonic development, tissue remodeling, tumor invasion, andinflammation. It activates the urokinase-type plasminogen activator,collagenases and several complement zymogens, such as C1 and C5. Itsrole in tissue remodeling and tumor invasion may be modulated by CSPG4.

The Protein Data Bank website provides the crystal structure of PLGsearchable by 1DDJ (Wang, X. et al., J.Mol.Biol., 2000, 295, 903-914);and 4DUR and 4DUU (Law, R. H. P., et al., Cell Rep., 2012, 1, 185-190).

Representative PLG Targeting Ligands are provided in FIG. 1 . AdditionalPLG Targeting Ligands are provided in, for example, J Med Chem 35:4297-305 (1992), J Med Chem 38: 1511-22 (1995), J Med Chem 56: 820-31(2013), U.S. Pat. Nos. 8,598,206, 8,921,319, J Med Chem 55: 1171-80(2012), Bioorg Med Chem Lett 12: 3183-6 (2002), Bioorg Med Chem 23:3696-704 (2015), Bioorg Med Chem Lett 13: 723-8 (2003), Bioorg Med ChemLett 7: 331-336 (1997), each of which is incorporated by referenceherein.

Plasminogen Activator Inhibitor-1 (PAI-1)

In some embodiments, the Target Protein is human plasminogen activatorinhibitor 1 (PAI-1) (UniProtKB—P05121 (PAI1_HUMAN)). PAI-I is a serineprotease inhibitor, and a primary inhibitor of tissue-type plasminogenactivator (PLAT) and urokinase-type plasminogen activator (PLAU). AsPLAT inhibitor, it is required for fibrinolysis down-regulation and isresponsible for the controlled degradation of blood clot. As PLAUinhibitor, it is involved in the regulation of cell adhesion andspreading, and acts as a regulator of cell migration, independently ofits role as protease inhibitor. Overexpression of PAI-I favorsangiogenesis, metastasis, and poor prognosis in tumors, including, butnot limited to, oral cancers and breast cancers.

The Protein Data Bank website provides the crystal structure of PAI-Isearchable by 3Q02 and 3Q03 (Jensen, J. K. et al., J Biol Chem., 2011,286, 29709-29717); 1B3K (Sharp, A. M. et al., Structure, 1999, 7,111-118); 1C5G (Tucker, H. M. et al., Nat Struct Biol., 1995, 2,442-445); 1DVM (Stout, T. J. et al., Biochemistry, 2000, 39, 8460-8469);and 3UT3 (Lin, Z. H. et al.,); as well as the crystal structure of PAI-Ibound to various compounds searchable by 4AQH (Fjellstrom, O. et al., JBiol Chem., 2013, 288, 873); 3R4L (Jankun, J. et al., Int J Mol Med.,2012, 29 61-64); 1A7C (Xue, Y., et al., Structure, 1998, 6, 627-636);1OCO (Zhou, A. et al., Nat Struct Biol., 2003, 10, 541); 6I8S (Vousden,K. A. et al., Sci Rep., 2019, 9, 1605-1605); 4G80 and 4G8R (Li, S. H. etal., Proc Natl Acad Sci USA, 2013, 110, E4941-E4949); 6GWQ, 6GWN and6GWP (Sillen, M. et al., J Thromb Haemost, 2019); and 4ICO (Hong, Z. B.et al.,).

Representative PAI-I Targeting Ligands are provided in FIG. 1 .Additional PAI-I Targeting Ligands are provided in, for example, J BiolChem 285: 7892-902 (2010), U.S. Pat. No. 9,120,744, Bioorg Med Chem Lett13: 3361-5 (2003), Bioorg Med Chem Lett 12: 1063-6 (2002), Bioorg MedChem Lett 13: 1705-8 (2003), Bioorg Med Chem Lett 11: 2589-92 (2001),U.S. Pat. No. 9,718,760, each of which is incorporated by referenceherein.

Placenta Growth Factor (PIGF)

In some embodiments, the Target Protein is human placental growth factor(PGF) (UniProtKB—P49763 (PLGF_HUMAN)). PGF is growth factor active inangiogenesis and endothelial cell growth, stimulating theirproliferation and migration. It binds to the receptor FLT1/VEGFR-1.Isoform PlGF-2 binds NRP1/neuropilin-1 and NRP2/neuropilin-2 in aheparin-dependent manner. PGF also promotes cell tumor growth, and hasbeen implicated in age-related macular degeneration (AMD) and choroidalneovascularization (CNV).

The Protein Data Bank website provides the crystal structure of PIGFsearchable by 1FZV (Iyer, S. et al., J Biol Chem., 2001, 276,12153-12161); as well as the crystal structure of PIGF bound to variouscompounds searchable by 1RV6 (Christinger, H. W., J Biol Chem., 2004,279, 10382-10388). Additionally, De Falco provides insight into thediscovery and biological activity of placenta growth factor (De Falco,Exp Mol Med., 2012, 44, 1-9).

Representative PGF Targeting Ligands are provided in FIG. 1 . AdditionalPGF Targeting Ligands are provided in, for example, J Med Chem 54:1256-65 (2011), J Nat Prod 76: 29-35 (2013), each of which isincorporated by reference herein.

Phospholipase A2, Group IB (PA21B)

In some embodiments, the Target Protein is human phospholipase A2, GroupIB (PA21B) (UniProtKB—P04054 (PA21B_HUMAN)). PA21B cleaves phospholipidspreferentially at the sn-2 position, liberating free fatty acids andlysophospholipids. PA21B has been implicated in a number of diseases,including cardiovascular diseases, atherosclerosis, immune disorders andcancer.

The Protein Data Bank website provides the crystal structure of PA21Bsearchable by 3FVJ and 3FVI (Pan, Y. H. et al., Biochim.Biophys.Acta.,2010, 1804, 1443-1448).

Representative PA21B Targeting Ligands are provided in FIG. 1 .Additional PA21B Targeting Ligands are provided in, for example, J MedChem 39: 3636-58 (1996), Chembiochem 4: 181-5 (2003), J Med Chem 39:5159-75 (1997), J Med Chem 51: 4708-14 (2008), each of which isincorporated by reference herein.

Phospholipase A2, Group IIA (PA2GA)

In some embodiments, the Target Protein is human phospholipase A2, GroupIIA (PA2GA) (UniProtKB—P04054 (PA21B_HUMAN)). PA2GA catalyzes thecalcium-dependent hydrolysis of the 2-acyl groups in3-sn-phosphoglycerides. It is thought to participate in the regulationof phospholipid metabolism in biomembranes including eicosanoidbiosynthesis. Independent of its catalytic activity, it also acts as aligand for integrins. PA2GA Induces cell proliferation in anintegrin-dependent manner. PA2GA has been implicated in a number ofdiseases, including cardiovascular diseases, atherosclerosis, immunedisorders, and cancer.

The Protein Data Bank website provides the crystal structure of PA2GAbound to various compounds searchable by 2ARM and 1SV3 (Singh, N. etal., Proteins, 2006, 64, 89-100); 5G3M and 5G3N (Giordanetto, F., et al.ACS Med Chem Lett., 2016, 7, 884); 1KQU (Jansford, K. A., et al.,Chembiochem., 2003, 4,181-185); and 1ZYX (Singh, N. et al.,).Additionally, Singh et al., provides insight into the crystal structureof the complexes of a group IIA phospholipase A2 with two naturalanti-inflammatory agents, anisic acid, and atropine reveal a similarmode of binding (Singh, N. et al., Proteins, 2006, 64(1):89-100); andKitadokoro et al also provides insight into the crystal structure ofhuman secretory phospholipase A2-IIA complex with the potent indolizineinhibitor 120-1032 (Kitadokoro, K. et al., J Biochem., 1998, 123(4),619-23).

Representative PA2GA Targeting Ligands are provided in FIG. 1 .Additional PA2GA Targeting Ligands are provided in, for example, J MedChem 48: 893-6 (2005), J Med Chem 39: 5159-75 (1997), each of which isincorporated by reference herein.

Factor B

In some embodiments, the Target Protein is human Complement factor B(UniProtKB-P00751 (CFAB_HUMAN)). Complement factor B, which is part ofthe alternate pathway of the complement system, is cleaved by factor Dinto 2 fragments: Ba and Bb. Bb, a serine protease, then combines withcomplement factor 3b to generate the C3 or C5 convertase. It has alsobeen implicated in proliferation and differentiation of preactivatedB-lymphocytes, rapid spreading of peripheral blood monocytes,stimulation of lymphocyte blastogenesis and lysis of erythrocytes. Bainhibits the proliferation of preactivated B-lymphocytes.

The Protein Data Bank website provides the crystal structure ofComplement Factor B searchable by 2OK5 (Milder, F. J., et al., NatStruct Mol Bio 2007, 14, 224-228); as well as the crystal structure ofComplement factor B bound to various compounds searchable by 6QSW, 6QSX,and 6RAV (Schubart, A., et al., Proc Natl Acad Sci 2019, 116,7926-7931); 6T8U, 6T8W, and 6T8V (Mainolfi, N., et al, J Med Chem 2020,63, 5697-5722); and 7JTN (Xu, X., et al., J Immunol 2021, 206,doi:10.4049/jimmunol.2001260).

Representative Complement Factor B Targeting Ligands are provided inFIG. 5 . Additional Complement Factor B Targeting Ligands are providedin, for example, U.S. Pat. No. 9,682,968B2, U.S. Pat. No. 9,475,806B2,U.S. Pat. No. 9,452,990B2, Proc Natl Acad Sci 116: 7926-7931 (2019), JMed Chem 52: 6042-6052 (2009), and J Med Chem 63: 5697-5722 (2020), eachof which is incorporated by reference herein.

In certain embodiments the Extracellular Targeting Ligand is selectedfrom:

each of which is optionally substituted with 1, 2, 3, or 4 substituentsindependently selected from R²¹.

In certain embodiments the Factor B Targeting Ligand is selected from aligand described in: Mainolfi, N. et. al. Discovery of 4-((2 S,4S)-4-Ethoxy-1-((5-Methoxy-7-Methyl-1H-Indol-4-Yl)Methyl)Piperidin-2-Yl)Benzoic Acid (LNPO23), a Factor BInhibitor Specifically Designed To Be Applicable to Treating a DiverseArray of Complement Mediated Diseases. J. Med. Chem. 2020, 63 (11),5697-5722; WO2020/016749; WO2018/005552; WO2013/192345; or WO2015009616.

In certain embodiments the factor B Targeting Ligand-linker is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom the following compounds or a bi- or tri-dentate version thereof:

Factor D

In some embodiments, the Target Protein is human Complement factor D(UniProtKB-P00746 (CFAD_HUMAN)). Factor D cleaves factor B when thelatter is complexed with factor C3b, activating the C3bbb complex, whichthen becomes the C3 convertase of the alternate pathway. Its function ishomologous to that of CIs in the classical pathway.

The Protein Data Bank website provides the crystal structure ofComplement factor D bound to various compounds searchable by 6FTZ, 6FUT,6FUH, 6FUG, 6FUJ, and 6FUI (Vulpetti, A., et al., ACS Med Chem Lett2018, 9, 490-495); 5TCA and 5TCC (Yang, C. Y., et al., ACS Med Chem Lett2016, 7, 1092-1096); 5MT4 (Vulpetti, A., et al., J Med Chem 2017, 60,1946-1958); 1DFP (Cole, L. B., et al., Acta Crystallogr D BiolCrystallogr 1997, 53, 143-150); 1DIC (Cole, L. B., et al., ActaCrystallogr D Biol Crystallogr 1998, 54, 711-717); 6QMR and 6QMT (Karki,R. G., et al., J Med Chem 2019, 62, 4656-4668).

Representative Complement factor D Targeting Ligands are provided inFIG. 6 . Additional Complement Factor D Targeting Ligands are providedin, for example, J Med Chem 60: 5717-5735 (2017), Nat Chem Biol 12:1105-1110 (2016), U.S. Pat. No. 9,598,446B2, U.S. Pat. No. 9,643,986B2,US patent U.S. Pat. No. 9,663,543B2 US patent U.S. Pat. No. 9,695,205B2,U.S. Pat. No. 9,732,103B2, U.S. Pat. No. 9,732,104B2, U.S. Pat. No.9,758,537B2, U.S. Pat. No. 9,796,741B2, U.S. Pat. No. 9,828,396B2, U.S.patent Ser. No. 10/000,516B2, U.S. patent Ser. No. 10/005,802B2, U.S.patent Ser. No. 10/011,612B2, U.S. patent Ser. No. 10/081,645B2, U.S.patent Ser. No. 10/087,203B2, U.S. patent Ser. No. 10/092,584B2, U.S.patent Ser. No. 10/100,072B2, U.S. patent Ser. No. 10/106,563B2, U.S.patent Ser. No. 10/138,225B2, U.S. patent Ser. No. 10/189,869B2, U.S.patent Ser. No. 10/253,053B2, U.S. patent Ser. No. 10/287,301B2, U.S.patent Ser. No. 10/301,336B2, U.S. patent Ser. No. 10/370,394B2, U.S.patent Ser. No. 10/385,097B2, U.S. patent Ser. No. 10/428,094B2, U.S.patent Ser. No. 10/428,095B2, U.S. patent Ser. No. 10/464,956B2, U.S.patent Ser. No. 10/550,140B2, U.S. patent Ser. No. 10/660,876B2, U.S.patent Ser. No. 10/662,175B2, U.S. patent Ser. No. 10/689,409B2, U.S.patent Ser. No. 10/807,952B2, U.S. patent Ser. No. 10/822,352B2, U.S.Pat. No. 9,464,081B2, and Haematologica 102: 466-475 (2017), each ofwhich is incorporated by reference herein.

In certain embodiments the Extracellular Targeting Ligand is selectedfrom:

wherein:

R^(21a), R^(21b), R^(21c), R^(21d), R^(21e), R^(21f), and R^(21g) areindependently at each occurrence selected from the group consisting ofhydrogen, alkyl, alkenyl, alkynyl, F, Cl, Br, I, hydroxyl, alkoxy,azide, amino, cyano, —NR⁶R⁷, —NR⁸SO₂R³, —NR'S(O)R³, haloalkyl,heteroalkyl, aryl, heteroaryl, heterocyclyl, —SR³, —C(O)OR³,—C(O)NR⁶NR⁷, —OR³, and heterocycle;

R²⁰¹, R²⁰², R²⁰²′, and R²⁰³ are independently selected from hydrogen,halogen, hydroxyl, nitro, cyano, amino, C₁-C₆alkyl, C₂-C₆alkenyl,C₁-C₆alkoxy, C₂-C₆alkynyl, C₂-C6alkanoyl, C₁-C₆thioalkyl,hydroxyC₁-C₆alkyl, aminoC₁-C₆alkyl, —C₀-C₄alkylNR⁹R¹⁰, —C(O)OR⁹,—OC(O)R⁹, —NR⁹C(O)R¹⁰, —C(O)NR⁹R¹⁰, —OC(O)NR⁹R¹⁰, —O(heteroaryl),—NR⁹C(O)OR¹⁰, C₁-C₂haloalkyl, -C₀-C₄alkyl(C3-C7cycloalkyl) and—O—C₀-C₄alkyl(C3-C7cycloalkyl), and C₁-C₂haloalkoxy, where R²⁰⁹ and R²¹⁰are independently chosen at each occurrence from hydrogen, C₁-C₆alkyl,and (C₃-C₇cycloalkyl)C₀-C₄alkyl;

or R²⁰² and R²⁰²′ may be taken together to form a 3- to 6-membered spiroring optionally substituted with 1 or more substituents independentlychosen from halogen, hydroxyl, cyano, —COOH, C₁-C₄alkyl (including inparticular methyl), C₂-C₄alkenyl, C₂-C₄alkynyl, C₁-C₄alkoxy,C₂-C4alkanoyl, hydroxyC₁-C₄alkyl, (mono— anddi-C₁-C₄alkylamino)C₀-C₄alkyl, -C₀-C₄alkyl(C₃-C₇cycloalkyl),—O—C₀-C₄alkyl(C₃-C₇cycloalkyl), C₁-C₂haloalkyl, and C₁-C₂haloalkoxy.

or R²⁰ and R²⁰² may be taken together to form a 3-membered carbocyclicring, optionally substituted with 1, 2, or 3 substituents selected fromR²¹.

or R²⁰¹ and R²⁰² may be taken together to form a 4- to 6-memberedcarbocyclic ring or a 4- to 6-membered heterocyclic ring containing 1 or2 heteroatoms independently chosen from N, O, and S, optionallysubstituted with 1, 2, or 3 substituents selected from R²¹.

R²⁰² and R²⁰³ may be taken together to form a 3- to 6-memberedcarbocyclic ring or a 3- to 6-membered heterocyclic ring optionallysubstituted with 1, 2, or 3 substituents selected from R²¹.

L¹⁰⁰ is selected from

wherein R²¹⁷ is hydrogen or C₁-C₆alkyl and R²¹⁸ and R²¹⁸′ areindependently chosen from hydrogen, halogen, 10 hydroxymethyl, andmethyl; and m is 0, 1, 2, or 3;

B¹⁰⁰ is a cycloalkyl, heterocycle group having 1, 2, 3, or 4 heteroatomsindependently selected from N, O, and S, a C₂-C₆alkenyl, C₂-C₆alkynylgroup,—(C₀-C₄alkyl)(aryl), —(C₀-C₄alkyl)(heteroaryl), or—(C₀-C₄alkyl)(biphenyl), each of which is optionally substituted with 1,2, 3, or 4 substituents independently selected from R²¹.

In certain embodiments the Extracellular Targeting Ligand is selectedfrom:

each of which is optionally substituted with 1, 2, 3, or 4 substituentsindependently selected from R²¹

In certain embodiments the Factor D Targeting Ligand is selected from aligand described in: U.S. Patent 9,796,74; U.S. Pat. No. 10,011,612;WO2018/160889; WO2019/195720; WO2019/057946; Karki, R. G. et al. Design,Synthesis, and Preclinical Characterization of Selective Factor DInhibitors Targeting the Alternative Complement Pathway. J. Med. Chem.2019, 62 (9), 4656-4668; or Belanger, D. B. et al.; WO2015/009977.

In certain embodiments the complement factor D targetingligand-linker-is selected from:

In certain embodiments the compound of the present invention is selectedfrom the following compounds or a bi- or tri-dentate version thereof:

Factor H

In some embodiments, the Target Protein is human complement factor H(UniProtKB-P08603 (CFAH_HUMAN)). Complement factor H is a glycoproteinthat plays an essential role in maintaining a well-balanced immuneresponse by modulating complement activation. Acts as a solubleinhibitor of complement, where its binding to self-markers such asglycan structures prevents complement activation and amplification oncell surfaces. Complement factor H accelerates the decay of thecomplement alternative pathway (AP) C₃ convertase C3bBb, thus preventinglocal formation of more C3b, the central player of the complementamplification loop. As a cofactor of the serine protease factor I, CFHalso regulates proteolytic degradation of already-deposited C3b. Inaddition, it mediates several cellular responses through interactionwith specific receptors. For example, CFH interacts with CR3/ITGAMreceptor and thereby mediates the adhesion of human neutrophils todifferent pathogens. In turn, these pathogens are phagocytosed anddestroyed.

The Protein Data Bank website provides the crystal structure of highlysimilar mutants of complement factor H searchable by 3KXV and 3KZJ(Bhattacharjee, A., et al., Mol Immunol 2010, 47, 1686-1691); as well asthe crystal structure of wild type complement factor H bound to variouscompounds searchable by 2UWN (Prosser, B. E., et al., J Exp Med 2007,204, 2277); 5WTB (Zhang, Y., et al., Biochem J 2017, 474, 1619-1631);5032 and 5035 (Xue, X., et al., Nat Struct Mol Biol 2017, 24, 643-651);40NT (Blaum, B. S., et al., Nat Chem Biol 2015, 11, 77-82); and 4ZH1(Blaum, B. S., et al., Glycobiology 2016, 26, 532-539).

Representative complement factor H Targeting Ligands are provided inFIG. 7 . Additional complement factor H Targeting Ligands are providedin, for example, J Immunol 182: 6394-6400 (2009), PLoS Pathogens 4:e1000250 (2008), PLoS Pathogens 6: e1001027 (2010), U.S. patent Ser. No.10/865,238B1, U.S. Pat. No. 8,962,795B2, US patent application20160317573A1, and US patent application 20190315842A1, each of which isincorporated by reference herein.

Complement Component 5 (C5)

In some embodiments, the Target Protein is human complement component 5(C5) (UniProtKB—P01031 (CO5_HUMAN)). Activation of C5 by a C5 convertaseinitiates the spontaneous assembly of the late complement components,C5-C9, into the membrane attack complex. C5b has a transient bindingsite for C6. The C5b-C6 complex is the foundation upon which the lyticcomplex is assembled.

The Protein Data Bank website provides the crystal structure ofComplement Component 5 searchable by 3CU7 (Fredslund, F., Nat Immunol2008, 9, 753-760); as well as the crystal structure of ComplementComponent 5 bound to various compound searchable by 5I5K(Schatz-Jakobsen, J. A., et al, J Immunol 2016, 197, 337-344); 3PVM and3PRX (Laursen, N. S., et al., EMBO J 2011, 30, 606-616); and 3KLS(Laursen, N. S., et al., Proc Natl Acad Sci 2010, 107, 3681-3686).

Representative Complement Component 5 Targeting Ligands are provided inFIG. 8 . Additional Complement Component 5 Targeting Ligands areprovided in, for example, J Immunol 197: 337-344 (2016), Ther AdvHematol 10: 1-11 (2019), BioDrugs 34: 149-158 (2020), Blood 135: 884-885(2020), US patent application 20170342139A1, and US patent application20200095307A1, each of which is incorporated by reference herein.

In certain embodiments the Extracellular Targeting Ligand is selectedfrom:

each of which is optionally substituted with 1, 2, 3, or 4 substituentsindependently selected from R²¹.

In certain embodiments the complement C5 Targeting Ligand is selectedfrom a ligand described in Jendza, K. et al. A Small-Molecule Inhibitorof C5 Complement Protein. Nat Chem Biol 2019, 15 (7), 666-668; or Zhang,M.; Yang, X.-Y.; Tang, W.; Groeneveld, T. W. L.; He, P.-L.; Zhu, F.-H.;Li, J.; Lu, W.; Blom, A. M.; Zuo, J.-P.; Nan, F.-J. Discovery andStructural Modification of 1-Phenyl-3-(1-Phenylethyl)Urea Derivatives asInhibitors of Complement. ACS Med. Chem. Lett. 2012, 3 (4), 317-321.

In certain embodiments the C5 Targeting Ligand is selected from:

5 Complement C1s

In certain embodiments the extracellular targeting ligand is a C1sTargeting Ligand.

In certain embodiments the complement C1s Targeting Ligand is selectedfrom a ligand described in WO2020/198062 or U.S. Pat. No. 6,683,055.

In certain embodiments the compound of the present invention is selectedfrom the 10 following compounds or a bi- or tri-dentate version thereof:

MASP

In certain embodiments the extracellular targeting ligand is a MASPTargeting Ligand.

In certain embodiments the MASP Targeting Ligand is selected from aligand described in Héja, D. et al. Monospecific Inhibitors Show ThatBoth Mannan-Binding Lectin-Associated Serine Protease-1 (MASP-1) and—2Are Essential for Lectin Pathway Activation and Reveal StructuralPlasticity of MASP-2. Journal of Biological Chemistry 2012, 287 (24),20290-20300; Dobó, J.; Kocsis, A.; Gil, P. Be on Target: Strategies ofTargeting Alternative and Lectin Pathway Components inComplement-Mediated Diseases. Front. Immunol. 2018, 9, 1851; or WO2014/144542.

In certain embodiments the MSAP-1 Targeting Ligand is SGMI-1 peptide,linked through the N- or C-terminus.

In certain embodiments the MSAP-1 Targeting Ligand is SGMI-2 peptide,linked through the N- or C-terminus.

In certain embodiments the MSAP-1 Targeting Ligand is TFMI-3 peptide,linked through the N- or C-terminus.

Factor XIa

In certain embodiments the extracellular targeting ligand is a factorXIa Targeting Ligand.

In certain embodiments the factor XIa Targeting Ligand is selected froma ligand described in: Lorthiois, E. et al. Structure-Based Design andPreclinical Characterization of Selective and Orally Bioavailable FactorXIa Inhibitors: Demonstrating the Power of an Integrated S1 ProteaseFamily Approach. J. Med. Chem. 2020, 63 (15), 8088-8113.

In certain embodiments the factor XIa Targeting Ligand is selected froma ligand described in: Quan, M. L. et al. Factor XIa Inhibitors as NewAnticoagulants. J. Med. Chem. 2018, 61 (17), 7425-7447.

In certain embodiments the factor XIa Targeting Ligand is selected froma ligand described in: Yang, W. et al. Discovery of a High Affinity,Orally Bioavailable Macrocyclic FXIa Inhibitor with AntithromboticActivity in Preclinical Species. J. Med. Chem. 2020, 63 (13), 7226-7242.

In certain embodiments the factor XIa Targeting Ligand-linker is:

In certain embodiments the compound of the present invention is selectedfrom the following compounds or a bi- or tri-dentate version thereof:

In certain embodiments the factor Xia Targeting Ligand is selected wherean anchor bond is placed at any suitable location with or withoutfunctionalization.

Additional Complement Extracellular Targeting Ligands

In certain embodiments the Extracellular Targeting Ligand is selectedfrom OMS721, Amy 101, APL2, ACH-4471, LNPO23, eculizumab, and avacopan.In other embodiments the 10 extracellular targeting ligand is selectedfrom C1-INH, rhucin, TP10, CAB-2, eculizumab, pexelizumab, ofatumumab,compstatin, PMX-53, and rhMBL. In other embodiments the extracellulartargeting ligand is selected from BCX1470, TP-20, mirococept, TNX-234,TNX-558, TA106, neutrazumab, anti-properdin, HuMax-CD38, ARC1905, andJPE-1375.

TNF-alpha

In certain embodiments the Extracellular Targeting Ligand is a TNF-alphaTargeting Ligand.

In certain embodiments the TNF-alpha Targeting Ligand is selected from aligand described in Dietrich, J. D. et al. Development of OrallyEfficacious Allosteric Inhibitors of TNFα via Fragment-Based DrugDesign. J. Med. Chem. 2021, 64 (1), 417-429.

In certain embodiments the TNF-alpha Targeting Ligand-linker is selectedfrom:

In certain embodiments the compound of the present invention is selectedfrom the following compounds or a bi- or tri-dentate version thereof:

Specific Extracellular Targeting Ligands

In certain embodiments the Extracellular Protein Targeting Ligand isOPT-3. OPT-3 has the following structure. It can bound to the linker inany available location using standard linking chemistry.

In certain embodiments OPT-3 is attached to the linker through theprimary amine of histidine as shown below.

OPT-Nil₂ has the structure shown below

In certain embodiments OPT-3 is attached to the linker through analkyne-azide click reaction. OPT-alkyne has the following structure.

In certain embodiments the Extracellular Protein Targeting Ligand isOT-2. OPT-2 has the following structure.

In certain embodiments OPT-2 is attached to the linker through theprimary amine of histidine as shown below.

In certain embodiments the Extracellular Protein Targeting Ligand isOPT-1. OPT-1 has the following structure.

In certain embodiments OPT-1 is attached to the linker through theprimary amine of histidine as shown below.

IV. Pharmaceutical Compositions and Dosage Forms for the ExtracellularDegrading Compounds of the Present Invention

A compound of the present invention or a pharmaceutically acceptablesalt, solvate or prodrug thereof as disclosed herein can be administeredas a neat chemical, but is more typically administered as apharmaceutical composition that includes an effective amount for a host,typically a human, in need of such treatment to treat a disordermediated by the target extracellular protein, as described herein orotherwise well-known for that extracellular protein.

The ASGPR-binding Extracellular Protein degraders of the presentinvention can be administered in any manner that allows the degrader tobind to the Extracellular Protein, typically in the blood stream, andcarry it to the ASGPR-bearing hepatocyte cells on the liver forendocytosis and degradation. As such, examples of methods to deliver thedegraders of the present invention include, but are not limited to,oral, intravenous, sublingual, subcutaneous, parenteral, buccal, rectal,intra-aortal, intracranial, subdermal, transdermal, controlled drugdelivery, intramuscular, or transnasal, or by other means, in dosageunit formulations containing one or more conventional pharmaceuticallyacceptable carriers, as appropriate. In certain embodiments, thedegrader is provided in a liquid dosage form, a solid dosage form, agel, particle, etc.

In certain embodiments the compound of the present invention isadministered subcutaneously. Typically, the compound will be formulatedin a liquid dosage form for subcutaneous injection, such as a bufferedsolution. Non-limiting examples of solutions for subcutaneous injectioninclude phosphate buffered solution and saline buffered solution. Incertain embodiments the solution is buffered with multiple salts.

In certain embodiments the compound of the present invention isadministered intravaneously. Typically, the compound will be formulatedin a liquid dosage form for intravaneous injection, such as a bufferedsolution. Non-limiting examples of solutions for intravaneous injectioninclude phosphate buffered solution and saline buffered solution. Incertain embodiments the solution is buffered with multiple salts.

Therefore, the disclosure provides pharmaceutical compositionscomprising an effective amount of degrading compound or itspharmaceutically acceptable salt together with at least onepharmaceutically acceptable carrier for any appropriate use thereof. Thepharmaceutical composition may contain a compound or salt as the onlyactive agent, or, in an alternative embodiment, the compound and atleast one additional active agent.

The term “pharmaceutically acceptable salt” as used herein refers to asalt of the described compound which is, within the scope of soundmedical judgment, suitable for administration to a host such as a humanwithout undue toxicity, irritation, allergic response, and the like,commensurate with a reasonable benefit/risk ratio, and effective for itsintended use. Thus, the term “pharmaceutically acceptable salt” refersto the relatively non-toxic, inorganic and organic acid addition saltsof the presently disclosed compounds. These salts can be prepared duringthe final isolation and purification of the compounds or by separatelyreacting the purified compound in its free form with a suitable organicor inorganic acid and then isolating the salt thus formed. Basiccompounds are capable of forming a wide variety of different salts withvarious inorganic and organic acids. Acid addition salts of the basiccompounds are prepared by contacting the free base form with asufficient amount of the desired acid to produce the salt in theconventional manner. The free base form can be regenerated by contactingthe salt form with a base and isolating the free base in theconventional manner. The free base forms may differ from theirrespective salt forms in certain physical properties such as solubilityin polar solvents.

Pharmaceutically acceptable base addition salts may be formed with ametal or amine, such as alkali and alkaline earth metal hydroxide, or anorganic amine. Examples of metals used as cations, include, but are notlimited to, sodium, potassium, magnesium, calcium, and the like.Examples of suitable amines include, but are not limited to,N,N′-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine,ethylenediamine, N-methylglucamine, and procaine. The base additionsalts of acidic compounds are prepared by contacting the free acid formwith a sufficient amount of the desired base to produce the salt in theconventional manner. The free acid form can be regenerated by contactingthe salt form with an acid and isolating the free acid in a conventionalmanner. The free acid forms may differ from their respective salt formssomewhat in certain physical properties such as solubility in polarsolvents.

Salts can be prepared from inorganic acids sulfate, pyrosulfate,bisulfate, sulfite, bisulfite, nitrate, phosphate,monohydrogenphosphate, dihydrogenphosphate, metaphosphate,pyrophosphate, chloride, bromide, iodide, nitric, phosphoric, sulfuric,hydrobromic, hydriodic, phosphorus, and the like. Representative saltsinclude the hydrobromide, hydrochloride, sulfate, bisulfate, nitrate,acetate, oxalate, valerate, oleate, palmitate, stearate, laurate,borate, benzoate, lactate, phosphate, tosylate, citrate, maleate,fumarate, succinate, tartrate, naphthylate mesylate, glucoheptonate,lactobionate, laurylsulphonate and isethionate salts, and the like.Salts can also be prepared from organic acids, such as aliphatic mono—and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic andaromatic sulfonic acids, etc. and the like. Representative salts includeacetate, propionate, caprylate, isobutyrate, oxalate, malonate,succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate,chlorobenzoate, methylbenzoate, dinitrobenzoate, phthalate,benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate,maleate, tartrate, methanesulfonate, and the like. Pharmaceuticallyacceptable salts can include cations based on the alkali and alkalineearth metals, such as sodium, lithium, potassium, calcium, magnesium andthe like, as well as non-toxic ammonium, quaternary ammonium, and aminecations including, but not limited to, ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. Also contemplated are the saltsof amino acids such as arginate, gluconate, galacturonate, and the like.See, for example, Berge et al., J. Pharm. Sci., 1977, 66, 1-19, which isincorporated herein by reference.

Any dosage form can be used that achieves the desired results. Incertain embodiments the pharmaceutical composition is in a dosage formthat contains from about 0.1 mg to about 1500 mg, from about 10 mg toabout 1000 mg, from about 100 mg to about 800 mg, or from about 200 mgto about 600 mg of the active compound and optionally from about 0.1 mgto about 1500 mg, from about 10 mg to about 1000 mg, from about 100 mgto about 800 mg, or from about 200 mg to about 600 mg of an additionalactive agent in a unit dosage form. Examples are dosage forms with atleast 0.1, 1, 5, 10, 25, 50, 100, 200, 250, 300, 400, 500, 600, 700, or750 mg of active compound, or its salt.

In certain embodiments the dose ranges from about 0.01-100 mg/kg ofpatient bodyweight, for example about 0.01 mg/kg, about 0.05 mg/kg,about 0.1 mg/kg, about 0.5 mg/kg, about 1 mg/kg, about 1.5 mg/kg, about2 mg/kg, about 2.5 mg/kg, about 3 mg/kg, about 3.5 mg/kg, about 4 mg/kg,about 4.5 mg/kg, about 5 mg/kg, about 10 mg/kg, about 15 mg/kg, about 20mg/kg, about 25 mg/kg, about 30 mg/kg, about 35 mg/kg, about 40 mg/kg,about 45 mg/kg, about 50 mg/kg, about 55 mg/kg, about 60 mg/kg, about 65mg/kg, about 70 mg/kg, about 75 mg/kg, about 80 mg/kg, about 85 mg/kg,about 90 mg/kg, about 95 mg/kg, or about 100 mg/kg.

In some embodiments, compounds disclosed herein or used as described areadministered once a day (QD), twice a day (BID), or three times a day(TID). In some embodiments, compounds disclosed herein or used asdescribed are administered at least once a day for at least 1 day, atleast 2 days, at least 3 days, at least 4 days, at least 5 days, atleast 6 days, at least 7 days, at least 8 days, at least 9 days, atleast 10 days, at least 11 days, at least 12 days, at least 13 days, atleast 14 days, at least 15 days, at least 16 days, at least 17 days, atleast 18 days, at least 19 days, at least 20 days, at least 21 days, atleast 22 days, at least 23 days, at least 24 days, at least 25 days, atleast 26 days, at least 27 days, at least 28 days, at least 29 days, atleast 30 days, at least 31 days, at least 35 days, at least 45 days, atleast 60 days, at least 75 days, at least 90 days, at least 120 days, atleast 150 days, at least 180 days, or longer.

In certain embodiments the compound of the present invention isadministered once a day, twice a day, three times a day, or four times aday.

The pharmaceutical composition may be formulated as any pharmaceuticallyuseful form, e.g., a pill, capsule, tablet, an injection or infusionsolution, a syrup, an inhalation formulation, a suppository, a buccal orsublingual formulation, a parenteral formulation, or in a medicaldevice. Some dosage forms, such as tablets and capsules, can besubdivided into suitably sized unit doses containing appropriatequantities of the active components, e.g., an effective amount toachieve the desired purpose.

Carriers include excipients and diluents and must be of sufficientlyhigh purity and sufficiently low toxicity to render them suitable foradministration to the patient being treated. The carrier can be inert orit can possess pharmaceutical benefits of its own. The amount of carrieremployed in conjunction with the compound is sufficient to provide apractical quantity of material for administration per unit dose of thecompound. If provided as in a liquid, it can be a solution or asuspension.

Representative carriers include phosphate buffered saline, water,solvent(s), diluents, pH modifying agents, preservatives, antioxidants,suspending agents, wetting agent, viscosity agents, tonicity agents,stabilizing agents, and combinations thereof. In some embodiments, thecarrier is an aqueous carrier. Examples of aqueous carries include, butare not limited to, an aqueous solution or suspension, such as saline,plasma, bone marrow aspirate, buffers, such as Hank's Buffered SaltSolution (HBSS), HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonicacid), Ringers buffer, ProVisc®, diluted ProVisc®, Provisc® diluted withPBS, Krebs buffer, Dulbecco's PBS, normal PBS, sodium hyaluronatesolution (HA, 5 mg/mL in PBS), citrate buffer, simulated body fluids,plasma platelet concentrate and tissue culture medium or an aqueoussolution or suspension comprising an organic solvent. Acceptablesolutions include, for example, water, Ringer's solution and isotonicsodium chloride solutions. The formulation may also be a sterilesolution, suspension, or emulsion in a non-toxic diluent or solvent suchas 1,3-butanediol.

Viscosity agents may be added to the pharmaceutical composition toincrease the viscosity of the composition as desired. Examples of usefulviscosity agents include, but are not limited to, hyaluronic acid,sodium hyaluronate, carbomers, polyacrylic acid, cellulosic derivatives,polycarbophil, polyvinylpyrrolidone, gelatin, dextin, polysaccharides,polyacrylamide, polyvinyl alcohol (including partially hydrolyzedpolyvinyl acetate), polyvinyl acetate, derivatives thereof and mixturesthereof.

Solutions, suspensions, or emulsions for administration may be bufferedwith an effective amount necessary to maintain a pH suitable for theselected administration. Suitable buffers are well known by thoseskilled in the art. Some examples of useful buffers are acetate, borate,carbonate, citrate, and phosphate buffers. Solutions, suspensions, oremulsions for topical, for example, ocular administration may alsocontain one or more tonicity agents to adjust the isotonic range of theformulation. Suitable tonicity agents are well known in the art. Someexamples include glycerin, mannitol, sorbitol, sodium chloride, andother electrolytes.

Classes of carriers include, but are not limited to binders, bufferingagents, coloring agents, diluents, disintegrants, emulsifiers,flavorants, glidents, lubricants, preservatives, stabilizers,surfactants, tableting agents, and wetting agents. Some carriers may belisted in more than one class, for example vegetable oil may be used asa lubricant in some formulations and a diluent in others. Exemplarypharmaceutically acceptable carriers include sugars, starches,celluloses, powdered tragacanth, malt, gelatin; talc, and vegetableoils. Optional active agents may be included in a pharmaceuticalcomposition, which do not substantially interfere with the activity ofthe compound of the present invention.

The pharmaceutical compositions/combinations can be formulated for oraladministration. These compositions can contain any amount of activecompound that achieves the desired result, for example between 0.1 and99 weight % (wt. %) of the compound and usually at least about 5 wt. %of the compound. Some embodiments contain from about 25 wt. % to about50 wt. % or from about 5 wt. % to about 75 wt. % of the compound.Enteric coated oral tablets may also be used to enhance bioavailabilityof the compounds for an oral route of administration.

Formulations suitable for rectal administration are typically presentedas unit dose suppositories. These may be prepared by admixing the activecompound with one or more conventional solid carriers, for example,cocoa butter, and then shaping the resulting mixture.

VII. Treatment of Diseases with the Disclosed ASGPR-bindingExtracellular Protein Degraders

The Target Proteins of the current invention may include, but are notlimited to, immunoglobulins, cytokines, chemokines, growth factors,coagulation factors, extracellular matrix proteins and proteins involvedin formation and/or degradation of the extracellular matrix, esterases,lipases, peptidases, convertases, among others. These proteins mediate arange of diseases that can be treated with an effective amount of thedisclosed ASGPR-binding Extracellular Protein Degraders describedherein.

Immunoglobulins

-   -   1) Immunoglobulin A (IgA) aberrant expression mediates a range        of autoimmune and immune-mediated disorders, including IgA        nephropathy (also known as Berger's disease), celiac disease,        Crohn's disease, Henoch-Sconiein purpura (HSP) (also known as        IgA vasculitis), liner IgA bullous dermatosis, IgA pemphigus,        dermatitis herpetiformis, inflammatory bowel disease (IBD),        Sjogren's syndrome, ankylosing spondylitis, alcoholic liver        cirrhosis, acquired immunodeficiency syndrome, IgA multiple        myeloma, α-chain disease, IgA monoclonal gammopathy, monoclonal        gammopathy of undetermined significance (MGUS), and linear IgA        bullous dermatosis, among others.    -   2) Immunoglobulin G (IgG) mediates a range of autoimmune,        infectious and metabolic diseases, including systemic        fibroinflammatory disease. In addition, overexpression of IgG4        is associated with IgG4-related diseases, which generally        include multiple organs, and disorders include type 1 autoimmune        pancreatitis, interstitial nephritis, Riedel's thyroiditis,        storiform fibrosis, Mikulicz's disease, Kuttner's tumor,        inflammatory pseudotumors (in various sites of the body),        mediastinal fibrosis, retroperitoneal fibrosis (Ormond's        disease), aortitis and periaortitis, proximal biliary        strictures, idiopathic hypocomplementic tubulointerstitial        nephritis, multifocal fibrosclerosis, pachymeningitis,        pancreatic enlargement, tumefactive lesions, pericarditis,        rheumatoid arthritis (RA), inflammatory bowel disease, multiple        sclerosis, myasthenic gravis, ankylosing spondylitis, primary        Sjogren's syndrome, psoriatic arthritis, and systemic lupus        erythematosus (SLE), sclerosing cholangitis, and IgG monoclonal        gammopathy, monoclonal gammopathy of undetermined significance        (MGUS), among others.    -   3) Immunoglobulin E (IgE)—IgE is a strong mediator of allergic        disease, including but not limited to, atopic asthma, allergic        rhinitis, atopic dermatitis, IgE-mediated food allergy,        IgE-mediated animal allergies, allergic conjunctivitis, allergic        urticaria, anaphylactic shock, nasal polyposis,        keratoconjunctivitis, mastocytosis, and eosinophilic        gastrointestinal disease, bullous pemphigoid, chemotherapy        induced hypersensitivity reaction, seasonal allergic rhinitis,        interstitial cystitis, eosinophilic esophagitis, angioedema,        acute interstitial nephritis, atopic eczema, eosinophilic        bronchitis, chronic obstructive pulmonary disease,        gastroenteritis, hyper-IgE syndrome (Job's Syndrome), IgE        monoclonal gammopathy, and monoclonal gammopathy of undetermined        significance (MGUS), among others.

Cytokines/Chemokines

-   -   1) TNF-α mediates a number of disorders, including but not        limited to rheumatoid arthritis, inflammatory bowel disease,        graft-vs-host disease, ankylosing spondylitis, psoriasis,        hidradenitis suppurativa, refractory asthma, systemic lupis        erthyematosus, diabetes, and the induction of cachexia.    -   2) IL-2 mediates host versus graft rejection in transplants and        autoimmune disorders, including, but not limited to, multiple        sclerosis, idiopathic arthritis, iritis, anterior uveitis, IL-2        induced hypotension, psoriasis, and other autoimmune disorders    -   3) IL-1 mediates a number of auto-inflammatory and autoimmune        disorders, including, but not limited to, Blau syndrome,        cryopyrin-associated periodic syndromes, familial Mediterranean        fever, Majeed syndrome; mevalonate kinase deficiency syndrome,        pyogenic arthritis-pyoderma gangrenosum-acne syndrome, tumor        necrosis factor receptor-associated periodic syndrome, Behget's        Disease, Sjogren's Syndrome, gout and chondrocalcinosis,        periodic fever, aphthous stomatitis, pharyngitis, and cervical        adenitis (or PFAPA) syndrome, rheumatoid arthritis, Type 2        diabetes mellitus, acute pericarditis, Chronic interstitial lung        diseases (ILDs), and Still's disease amongst others.    -   4) IFN-γ mediates a wide range of autoimmune disorders,        including, but not limited to rheumatoid arthritis, multiple        sclerosis (MS), corneal transplant rejection, and various        autoimmune skin diseases such as psoriasis, alopecia areata,        vitiligo, acne vulgaris, and others.    -   5) IL-21 mediates a number of autoimmune disorders, including        Sjogren's syndrome, systemic lupus erythematosus, type 1        diabetes, multiple sclerosis, rheumatoid arthritis, and        inflammatory bowel disease.    -   6) IL-22 mediates a number of autoimmune disorders, including,        but not limited to, graft versus host disease (GVHD), psoriasis,        rheumatoid arthritis, atopic dermatitis, and asthma.    -   7) IL-10 has been implicated in tumor survival and protection        against cytotoxic chemotherapeutic drugs.    -   8) IL-5 has been implicated in a number of allergic disorders,        including, but not limited to, asthma, nasal polyposis, atopic        dermatitis, eosinophilic esophagitis, hypereosinophilic        syndrome, and Churg-Strauss syndrome.    -   9) IL-6 has been implicated in a number of inflammatory diseases        and cancers, including, but not limited to, Castleman's disease,        metastatic castration-associated prostate cancer, renal cell        carcinoma, large-cell lung carcinoma, ovarian cancer, rheumatoid        arthritis, asthma.    -   10)IL-8 has been implicated in the promotion of tumor        progression, immune escape, epithelial-mesenchymal transition,        and recruitment of myeloid-derived suppressor cells. Studies        have demonstrated that high serum IL-8 levels correlate with        poor prognosis in many malignant tumors. Preclinical studies        have shown that IL-8 blockade may reduce mesenchymal features in        tumor cells, making them less resistant to treatment.    -   11)C-C motif chemokine ligand 2 (CCL2) has been implicated in        the recruitment of monocytes into the arterial wall during the        disease process of atherosclerosis.    -   12)Macrophage Migration Inhibitory Factor (MIF) is a mediator of        tumor progression; systemic inflammation; atherosclerosis;        rheumatoid arthritis; and systemic lupus erythematosus, among        others.

Growth Factors

-   -   1) Fibroblast Growth Factor 1 (FGF1) can induce angiogenesis.        FGF1 has been implicated in oncogenesis, cancer cell        proliferation, resistance to anticancer therapies, and        neoangiogenesis.    -   2) Fibroblast Growth Factor 2 (FGF2) has been implicated in        oncogenesis, cancer cell proliferation, resistance to anticancer        therapies, and neoangiogenesis.    -   3) Vascular Epithelial Growth Factor (VEGF-A) has been        implicated in the vascularization and angiogenesis of tumors.    -   4) Transforming Growth Factor-(31 (TGF-β1) expression in the        tumor microenvironment has been associated with a poor        prognosis, and is implicated in TGF-β1 mediated tumor        suppression via T-cell exclusion. TGF-β1 expression has also        been implicated in hematological malignancies and fibrosis.    -   5) Transforming Growth Factor-02 (TGF-β2) expression in the        tumor microenvironment has been associated with a poor        prognosis, and is implicated in TGF-β2 mediated tumor        suppression via T-cell exclusion. TGF-β2 expression has also        been implicated in hematological malignancies and fibrosis.    -   6) Placental Growth Factor (PGF) promotes cell tumor growth, and        has been implicated in age-related macular degeneration (AMD)        and choroidal neovascularization (CNV).

Esterase

-   -   1) Cholinesterase has been implicated in cognitive disorders        such as dementia and Alzheimer's disease.

Coagulation Factors

-   -   1) Carboxypeptidase B2 has been implicated and targeted to        inhibit thrombosis.    -   2) Coagulation Factor Xa is a mediator in the development of        deep vein thrombosis and acute pulmonary embolism, and the risk        of stroke and embolism in people with nonvalvular atrial        fibrillation.    -   3) Coagulation Factor XI is a mediator in the development of        deep vein thrombosis and acute pulmonary embolism, and the risk        of stroke and embolism in people with nonvalvular atrial        fibrillation.    -   4) Coagulation Factor XII has been implicated in the development        of deep vein thrombosis and acute pulmonary embolism, and the        risk of stroke and embolism in people with nonvalvular atrial        fibrillation.    -   5) Coagulation Factor XIII has been implicated in the        development of deep vein thrombosis and acute pulmonary        embolism, and the risk of stroke and embolism in people with        nonvalvular atrial fibrillation.    -   6) Prothrombin is involved in blood clot formation and arterial        and venous thrombosis, and thromboembolism associated with        atrial fibrillation.    -   7) Coagulation Factor VII is involved in blood clot formation        and arterial and venous thrombosis, and thromboembolism        associated with atrial fibrillation.    -   8) Coagulation Factor IX is involved in blood clot formation and        arterial and venous thrombosis, and thromboembolism associated        with atrial fibrillation.

Extracellular Matrix Proteins

-   -   1) Neutrophil Elastase—Neutrophil elastase has been implicated        in a number of disorders, including lung disease, chronic        obstructive pulmonary disease, pneumonia, respiratory distress,        and acute lung injury (ALI), and cystic fibrosis, as well as        chronic kidney disease.    -   2) Fibronectin-1-Interfering with FN polymerization may        attenuate myofibroblasts and fibrosis and improve cardiac        function after ischemia/reperfusion (I/R) injury.    -   3) Thrombospondin-1- TSP-1 has been implicated in a number of        diseases, including in promoting certain cancers such as breast        cancer, prostate cancer, melanoma, SCLC, osteosarcoma, cutaneous        squamous cell carcinoma, oral squamous cell carcinoma, papillary        thyroid carcinoma, thyroid cancer, medulloblastoma, and fibrotic        disorders such as diabetes, liver fibrosis, and in multiple        myeloma.    -   4) Urokinase-type Plasminogen Activator (UPA)—UPA has been        implicated in vascular diseases and cancer progression. Elevated        expression levels of urokinase and several other components of        the plasminogen activation system are found to be correlated        with tumor malignancy.    -   5) Plasminogen Activator, Tissue Type (TPA)—PLA has been shown        activated in various cancers including oral malignancy.    -   6) Plasminogen (PLG)—PLG has been implicated in tumor invasion        and inflammation.    -   7) Plasminogen Activator Inhibitor-1 (PAI-1)—PAI-1 has been        implicated in angiogenesis, metastasis, and poor prognosis in        tumors, including, but not limited to, oral cancers and breast        cancers.

Peptidase

-   -   1) Kallikrein-1-Kallikrein has been implicated in adverse        reactions in hereditary angioedema (HAE).    -   2) Plasma Kallikrein—Plasma kallikrein has been implicated in        retinal dysfunction, the development of diabetic macular edema        and hereditary angioedema (HAE).    -   3) Matrix Metallopeptidase—1-MMP-1 has been implicated in        cardiovascular disease, development of fibrosis, and growth of        certain cancers such as bladder cancer.    -   4) Phospholipase A2, Group IIA (PA2GA)—PA2GA has been implicated        in a number of diseases, including cardiovascular diseases,        atherosclerosis, immune disorders, and cancer.

Lipase

-   -   1) Lipoprotein Lipase—Lipoprotein lipase has been implicated in        the development of cardiovascular disease and obesity.    -   2) Phospholipase A2, Group IB (PA21B)—PA21B has been implicated        in a number of diseases, including cardiovascular diseases,        atherosclerosis, immune disorders and cancer.

Convertase

-   -   1) Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK-9)—PCSK-9        has been implicated in high blood cholesterol and the        development of cardiovascular disease.

Certain extracellular protein targets include but are not limited to:SAA (serum amyloid A), amyloid light chains, antibodies to Kiebsielladipeptidase protein; Ig antibodies to anionic phospholipids andbeta-2-glycoprotein-I; IL-13; MIF; Transthyretin (misfolded), IgGautoantibodies to thyroid peroxidase, thyroglobulin and TSH receptors;TNF-α; Protein arginine deiminase (PAD, PAD4); antibodies tocitrullinated protein antibody (ACPA); anti-DNA antibodies; IL-17; LysylOxidase 2 (L.OXL2); IL-18; Blys; B cell activating factor (BAFF); CD40(soluble); CXCL12; soluble PSMA; matrix metalloproteinase IX (M-NMP-9);hormone-sensitive lipase; lipoprotein-associated phospholipase A2;Factor Xa; DPP4; thrombin; PCSK9; ApoB-100; Complement component C3b;PKK (pre-kallikrein); Factor XI; PF4; Anti-vWF antibodies;anticardiolipin antibodies and lupus anticoagulant; FGF23 (fibroblastgrowth factor 23); Plasminogen activator inhibitor type I (PAI-1);Myeloperoxidase (MPO) extracellular; Myostatin; Beta2-m; suPAR (solubleurokinase plasminogen activator receptor); anti-ganglioside IgG; anyloidbeta; Tau; CJD-associate prion; anti-ganglioside IgG; HTT;anti-ganglioside IgG; synuclein; elastase; PABA (protective antigen ofBacillus anthracis); edema factor; Botulinum toxin; C. difficile toxinB; hemolysin; tetanus toxin,IL-2; growth hormone and ACTH.

VIII Exemplary Methods of Treatment of Diseases Mediated byExtracellular Proteins

The present invention can be used to treat any disorder that is mediatedby the selected target disease-mediating extracellular protein.Nonlimiting examples of indications include autoimmune, other immunedysfunctions, complement mediated disorders, abnormal cellularproliferation, cancer, tumors, hematology-related disorders, renaldisorders and liver disorders.

In certain embodiments, the degrader or its salt or composition asdescribed herein is used in the treatment of an autoimmune disorder. Insome aspects, the extracellular protein is an Ig, such as IgA or IgG.IgG degradation can treat for example, thyroid eye disease, myastheniagravis, chronic inflammatory demyelinating polyneuropathy, and warmautoimmune hemolytic anemia.

Non-limiting examples of autoimmune disorders include: lupus, allograftrejection, autoimmune thyroid diseases (such as Graves' disease andHashimoto's thyroiditis), autoimmune uveoretinitis, giant cellarteritis, inflammatory bowel diseases (including Crohn's disease,ulcerative colitis, regional enteritis, granulomatous enteritis, distalileitis, regional ileitis, and terminal ileitis), diabetes, multiplesclerosis, pernicious anemia, psoriasis, rheumatoid arthritis,sarcoidosis, and scleroderma.

In an embodiment, the degrader or its salt or composition as describedherein is used in the treatment of lupus. Non-limiting examples of lupusinclude lupus erythematosus, cutaneous lupus, discoid lupuserythematosus, chilblain lupus erythematosus, or lupuserythematosus-lichen planus overlap syndrome. Lupus erythematosus is ageneral category of disease that includes both systemic and cutaneousdisorders. The systemic form of the disease can have cutaneous as wellas systemic manifestations. However, there are also forms of the diseasethat are only cutaneous without systemic involvement. For example, SLEis an inflammatory disorder of unknown etiology that occurspredominantly in women, and is characterized by articular symptoms,butterfly erythema, recurrent pleurisy, pericarditis, generalizedadenopathy, splenomegaly, as well as CNS involvement and progressiverenal failure. The sera of most patients (over 98%) contain antinuclearantibodies, including anti-DNA antibodies. High titers of anti-DNAantibodies are essentially specific for SLE. Conventional treatment forthis disease has been the administration of corticosteroids orimmunosuppressants.

There are three forms of cutaneous lupus: chronic cutaneous lupus (alsoknown as discoid lupus erythematosus or DLE), subacute cutaneous lupus,and acute cutaneous lupus. DLE is a disfiguring chronic disorderprimarily affecting the skin with sharply circumscribed macules andplaques that display erythema, follicular plugging, scales,telangiectasia and atrophy. The condition is often precipitated by sunexposure, and the early lesions are erythematous, round scaling papulesthat are 5 to 10 mm in diameter and display follicular plugging. DLElesions appear most commonly on the cheeks, nose, scalp, and ears, butthey may also be generalized over the upper portion of the trunk,extensor surfaces of the extremities, and on the mucous membranes of themouth. If left untreated, the central lesion atrophies and leaves ascar. Unlike SLE, antibodies against double-stranded DNA (e.g.,DNA-binding test) are almost invariably absent in DLE.

Multiple Sclerosis is an autoimmune demyelinating disorder that isbelieved to be T lymphocyte dependent. MS generally exhibits arelapsing-remitting course or a chronic progressive course. The etiologyof MS is unknown, however, viral infections, genetic predisposition,environment, and autoimmunity all appear to contribute to the disorder.Lesions in MS patients contain infiltrates of predominantly T lymphocytemediated microglial cells and infiltrating macrophages. CD4+Tlymphocytes are the predominant cell type present at these lesions. Thehallmark of the MS lesion is plaque, an area of demyelination sharplydemarcated from the usual white matter seen in MRI scans. Histologicalappearance of MS plaques varies with different stages of the disease. Inactive lesions, the blood-brain barrier is damaged, thereby permittingextravasation of serum proteins into extracellular spaces. Inflammatorycells can be seen in perivascular cuffs and throughout white matter.CD4+ T-cells, especially Th1, accumulate around postcapillary venules atthe edge of the plaque and are also scattered in the white matter.

In active lesions, up-regulation of adhesion molecules and markers oflymphocyte and monocyte activation, such as IL2-R and CD26 have alsobeen observed. Demyelination in active lesions is not accompanied bydestruction of oligodendrocytes. In contrast, during chronic phases ofthe disease, lesions are characterized by a loss of oligodendrocytes andhence, the presence of myelin oligodendrocyte glycoprotein (MOG)antibodies in the blood.

Diabetes can refer to either type 1 or type 2 diabetes. In someembodiments the degrader or its salt or composition as described hereinis provided at an effective dose to treat a patient with type 1diabetes. In one aspect the degrader or its salt or composition asdescribed herein is provided at an effective dose to treat a patientwith type 2 diabetes.

Type 1 diabetes is an autoimmune disease. An autoimmune disease resultswhen the body's system for fighting infection (the immune system) turnsagainst a part of the body. The pancreas then produces little or noinsulin.

As examples, the degrader or its salt or composition as described hereinis useful for treating or preventing a disorder selected from autoimmuneoophoritis, endometriosis, autoimmune orchitis, Ord's thyroiditis,autoimmune enteropathy, coeliac disease, Hashimoto's encephalopathy,antiphospholipid syndrome (APLS) (Hughes syndrome), aplastic anemia,autoimmune lymphoproliferative syndrome (Canale-Smith syndrome),autoimmune neutropenia, Evans syndrome, pernicious anemia, pure red cellaplasia, thrombocytopenia, adipose dolorosa (Dercum's disease), adultonset Still's disease, ankylosing spondylitis, CREST syndrome,drug-induced lupus, eosinophilic fasciitis (Shulman's syndrome), Feltysyndrome, IgG4-related disease, mixed connective tissue disease (MCTD),palindromic rheumatism (Hench-Rosenberg syndrome), Parry-Rombergsyndrome, Parsonage-Turner syndrome, relapsing polychondritis(Meyenburg-Altherr-Uehlinger syndrome), retroperitonial fibrosis,rheumatic fever, Schnitzler syndrome, fibromyalgia, neuromyotonia(Isaac's disease), paraneoplastic degeneration, autoimmune inner eardisease, Meniere's disease, interstitial cystitis, autoimmunepancreatitis, zika virus-related disorders, chikungunya virus-relateddisorders, subacute bacterial endocarditis (SBE), IgA nephropathy, IgAvasculitis, polymyalgia rheumatic, rheumatoid vasculitis, alopeciaareata, autoimmune progesterone dermatitis, dermatitis herpetiformis,erythema nodosum, gestational pemphigoid, hidradenitis suppurativa,lichen sclerosus, linear IgA disease (LAD), morphea, myositis,pityriasis lichenoides et varioliformis acuta, vitiligo post-myocardialinfarction syndrome (Dressler's syndrome), post-pericardiotomy syndrome,autoimmune retinopathy, Cogan syndrome, Graves opthalmopathy, ligneousconjunctivitis, Mooren's ulcer, opsoclonus myoclonus syndrome, opticneuritis, retinocochleocerebral vasculopathy (Susac's syndrome),sympathetic opthalmia, Tolosa-Hunt syndrome, interstitial lung disease,antisynthetase syndrome, Addison's disease, autoimmune polyendocrinesyndrome (APS) type I, autoimmune polyendocrine syndrome (APS) type II,autoimmune polyendocrine syndrome (APS) type III, disseminated sclerosis(multiple sclerosis, pattern II), rapidly progressing glomerulonephritis(RPGN), juvenile rheumatoid arthritis, enthesitis-related arthritis,reactive arthritis (Reiter's syndrome), autoimmune hepatitis or lupoidhepatitis, primary biliary cirrhosis (PBS), primary sclerosingcholangitis, microscopic colitis, latent lupus (undifferentiatedconnective tissue disease (UCTD)), acute disseminated encephalomyelitis(ADEM), acute motor axonal neuropathy, anti-n-methyl-D-aspartatereceptor encephalitis, Balo concentric sclerosis (Schilders disease),Bickerstaff's encephalitis, chronic inflammatory demyelinatingpolyneuropathy, idiopathic inflammatory demyelinating disease,Lambert-Eaton mysathenic syndrome, Oshtoran syndrome, pediatricautoimmune neuropsychiatric disorder associated with streptococcus(PANDAS), progressive inflammatory neuropathy, restless leg syndrome,stiff person syndrome, Sydenhem syndrome, transverse myelitis, lupusvasculitis, leukocytoclastic vasculitis, Microscopic Polyangiitis,polymyositis or ischemic-reperfusion injury of the eye.

In certain aspects, an effective amount of the degrader or its salt orcomposition as described herein is used to treat a medical disordermediated by the Targeted Extracellular Protein.

For example, when the Targeted Extracellular Protein is a complementprotein, for example complement factor B, factor D, factor H, C1s, C3,or C5, then the medical disorder to be treated may be an inflammatory orimmune condition, a disorder mediated by the complement cascade(including a dysfunctional cascade), or alternative complementpathway-related disorder, a disorder or abnormality of a cell thatadversely affects the ability of the cell to engage in or respond tonormal complement activity, or an undesired complement-mediated responseto a medical treatment, such as surgery or other medical procedure or apharmaceutical or biopharmaceutical drug administration, a bloodtransfusion, or other allogenic tissue or fluid administration.

In some aspects, the disorder treated by the degrader or its salt orcomposition as described herein is selected from fatty liver andconditions stemming from fatty liver, such as nonalcoholicsteatohepatitis (NASH), liver inflammation, cirrhosis and liver failure.

In other embodiments, the degrader or its salt or composition asdescribed herein is used to modulate an immune response prior to orduring surgery or other medical procedure. Non-limiting examples are theuse in connection with acute or chronic graft versus host disease, whichis a common complication as a result of allogeneic tissue transplant,and can also occur as a result of a blood transfusion.

In certain embodiments, the present invention provides a method oftreating or preventing dermatomyositis by administering to a subject inneed thereof an effective amount of the degrader or its salt orcomposition as described herein.

In certain embodiments, the present invention provides a method oftreating or preventing amyotrophic lateral sclerosis by administering toa subject in need thereof an effective amount of the degrader or itssalt or composition as described herein.

In certain embodiments, the present invention provides a method oftreating or preventing abdominal aortic aneurysm, hemodialysiscomplications, hemolytic anemia, or hemodialysis by administering to asubject in need thereof an effective amount of the degrader or its saltor composition as described herein.

In certain embodiments, a method is provided for the treatment orprevention of cytokine or inflammatory reactions in response to theadministration of pharmaceutical or biotherapeutic (e.g. CAR T-celltherapy or monoclonal antibody therapy) in a host by administering aneffective amount of the degrader or its salt or composition as describedherein. Various types of cytokine or inflammatory reactions may occur inresponse to a number of factors, such as the administrations ofbiotherapeutics. In one aspect, the cytokine or inflammatory reaction iscytokine release syndrome. In one embodiment, the cytokine orinflammatory reaction is tumor lysis syndrome (which also leads tocytokine release). Symptoms of cytokine release syndrome range fromfever, headache, and skin rashes to bronchospasm, hypotension and evencardiac arrest. Severe cytokine release syndrome is described ascytokine storm, and can be fatal.

In another embodiment, the disorder is episcleritis, idiopathicepiscleritis, anterior episcleritis, or posterior episcleritis. In oneembodiment, the disorder is idiopathic anterior uveitis, HLA-B27 relateduveitis, herpetic keratouveitis, Posner Schlossman syndrome, Fuch'sheterochromic iridocyclitis, or cytomegalovirus anterior uveitis.

In another embodiment, the present invention provides a method oftreating or preventing a C3 glomurenopathy by administering to a subjectin need thereof an effective amount of the degrader or its salt orcomposition as described herein. In one embodiment, the disorder isselected from dense deposit disease (DDD) and C₃ glomerulonephritis(C3GN).

In yet another embodiment, the present invention provides a method oftreating or preventing a IC-MPGN by administering to a subject in needthereof an effective amount of the degrader or its salt or compositionas described herein.

In a further embodiment, the present invention provides a method oftreating or preventing a paroxysmal nocturnal hemoglobinuria (PNH) byadministering to a subject in need thereof an effective amount of thedegrader or its salt or composition as described herein.

In another embodiment, the present invention provides a method oftreating or preventing age-related macular degeneration (AMD) byadministering to a subject in need thereof an effective amount of thedegrader or its salt or composition as described herein.

In one embodiment, the present invention provides a method of treatingor preventing rheumatoid arthritis by administering to a subject in needthereof an effective amount of the degrader or its salt or compositionas described herein.

In one embodiment, the present invention provides a method of treatingor preventing multiple sclerosis by administering to a subject in needthereof an effective amount of the degrader or its salt or compositionas described herein.

In one embodiment, the present invention provides a method of treatingor preventing myasthenia gravis by administering to a subject in needthereof an effective amount of the degrader or its salt or compositionas described herein.

In one embodiment, the present invention provides a method of treatingor preventing atypical hemolytic uremic syndrome (aHUS) by administeringto a subject in need thereof an effective amount of the degrader or itssalt or composition as described herein.

In one embodiment, the present invention provides a method of treatingor preventing neuromyelitis optica (NMO) by administering to a subjectin need thereof an effective amount of the degrader or its salt orcomposition as described herein.

In yet other embodiments, the present invention provides a method oftreating or preventing a disorder as described below by administering toa subject in need thereof an effective amount of the degrader or itssalt or composition as described herein, including: for example:vitritis, sarcoidosis, syphilis, tuberculosis, or Lyme disease; retinalvasculitis, Eales disease, tuberculosis, syphilis, or toxoplasmosis;neuroretinitis, viral retinitis, or acute retinal necrosis; varicellazoster virus, herpes simplex virus, cytomegalovirus, Epstein-Barr virus,lichen planus, or Dengue-associated disease (e.g., hemorraghic DengueFever); Masquerade syndrome, contact dermatitis, trauma inducedinflammation, UVB induced inflammation, eczema, granuloma annulare, oracne.

In additional embodiments, the disorder is selected from: acutemyocardial infarction, aneurysm, cardiopulmonary bypass, dilatedcardiomyopathy, complement activation during cardiopulmonary bypassoperations, coronary artery disease, restenosis following stentplacement, or percutaneous transluminal coronary angioplasty (PTCA);antibody-mediated transplant rejection, anaphylactic shock, anaphylaxis,allogenic transplant, humoral and vascular transplant rejection, graftdysfunction, graft-versus-host disease, Graves' disease, adverse drugreactions, or chronic graft vasculopathy; allergic bronchopulmonaryaspergillosis, allergic neuritis, drug allergy, radiation-induced lunginjury, eosinophilic pneumonia, radiographic contrast media allergy,bronchiolitis obliterans, or interstitial pneumonia;parkinsonism-dementia complex, sporadic frontotemporal dementia,frontotemporal dementia with Parkinsonism linked to chromosome 17,frontotemporal lobar degeneration, tangle only dementia, cerebralamyloid angiopathy, cerebrovascular disorder, certain forms offrontotemporal dementia, chronic traumatic encephalopathy (CTE), PD withdementia (PDD), argyrophilic grain dementia, dementia pugilistica,dementia with Lewy Bodies (DLB), or multi-infarct dementia;Creutzfeldt-Jakob disease, Huntington's disease, multifocal motorneuropathy (MMN), prion protein cerebral amyloid angiopathy,polymyositis, postencephalitic parkinsonism, subacute sclerosingpanencephalitis, non-Guamanian motor neuron disease with neurofibrillarytangles, neural regeneration, or diffuse neurofibrillary tangles withcalcification.

In further embodiments, the disorder is selected from: atopicdermatitis, dermatitis, dermatomyositis bullous pemphigoid, scleroderma,sclerodermatomyositis, psoriatic arthritis, pemphigus vulgaris, Discoidlupus erythematosus, cutaneous lupus, chilblain lupus erythematosus, orlupus erythematosus-lichen planus overlap syndrome; cryoglobulinemicvasculitis, mesenteric/enteric vascular disorder, peripheral vasculardisorder, antineutrophil cytoplasm antibody (ANCA)-associated vasculitis(AAV), TL-2 induced vascular leakage syndrome, or immune complexvasculitis;angioedema, low platelets (HELLP) syndrome, sickle celldisease, platelet refractoriness, red cell casts, or typical orinfectious hemolytic uremic syndrome (tHUS); hematuria, hemorrhagicshock, drug-induced thrombocytopenia, autoimmune hemolytic anemia(AIHA), azotemia, blood vessel and/or lymph vessel inflammation,rotational atherectomy, or delayed hemolytic transfusion reaction;British type amyloid angiopathy, Buerger's disease, bullous pemphigoid,Clq nephropathy, cancer, or catastrophic antiphospholipid syndrome.

In other embodiments, the disorder is selected from: wet (exudative)AMD, dry (non-exudative) AMD, chorioretinal degeneration, choroidalneovascularization (CNV), choroiditis, loss of RPE function, loss ofvision (including loss of visual acuity or visual field), loss of visionfrom AMD, retinal damage in response to light exposure, retinaldegeneration, retinal detachment, retinal dysfunction, retinalneovascularization (RNV), retinopathy of prematurity, pathologicalmyopia, or RPE degeneration; pseudophakic bullous keratopathy,symptomatic macular degeneration related disorder, optic nervedegeneration, photoreceptor degeneration, cone degeneration, loss ofphotoreceptor cells, pars planitis, scleritis, proliferativevitreoretinopathy, or formation of ocular drusen; chronic urticaria,Churg-Strauss syndrome, cold agglutinin disease (CAD), corticobasaldegeneration (CBD), cryoglobulinemia, cyclitis, damage of the Bruch'smembrane, Degos disease, diabetic angiopathy, elevated liver enzymes,endotoxemia, epidermolysis bullosa, or epidermolysis bullosa acquisita;essential mixed cryoglobulinemia, excessive blood urea nitrogen-BUN,focal segmental glomerulosclerosis, Gerstmann-Straussler-Scheinkerdisease, giant cell arteritis, gout, Hallervorden-Spatz disease,Hashimoto's thyroiditis, Henoch-Schonlein purpura nephritis, or abnormalurinary sediments; hepatitis, hepatitis A, hepatitis B, hepatitis C orhuman immunodeficiency virus (HIV), a viral infection more generally,for example selected from Flaviviridae, Retroviruses, Coronaviridae,Poxviridae, Adenoviridae, Herpesviridae, Caliciviridae, Reoviridae,Picornaviridae, Togaviridae, Orthomyxoviridae, Rhabdoviridae, orHepadnaviridae; Neisseria meningitidis, shiga toxin E. coli-relatedhemolytic uremic syndrome (STEC-HUS), hemolytic uremic syndrome (HUS);Streptococcus, or poststreptococcal glomerulonephritis.

In further embodiments, the disorder is selected from: hyperlipidemia,hypertension, hypoalbuminemia, hypobolemic shock, hypocomplementemicurticarial vasculitis syndrome, hypophosphastasis, hypovolemic shock,idiopathic pneumonia syndrome, or idiopathic pulmonary fibrosis;inclusion body myositis, intestinal ischemia, iridocyclitis, iritis,juvenile chronic arthritis, Kawasaki's disease (arteritis), orlipiduria; membranoproliferative glomerulonephritis (MPGN) I,microscopic polyangiitis, mixed cryoglobulinemia, molybdenum cofactordeficiency (MoCD) type A, pancreatitis, panniculitis, Pick's disease,polyarteritis nodosa (PAN), progressive subcortical gliosis,proteinuria, reduced glomerular filtration rate (GFR), or renovasculardisorder; multiple organ failure, multiple system atrophy (MSA),myotonic dystrophy, Niemann-Pick disease type C, chronic demyelinatingdiseases, or progressive supranuclear palsy; spinal cord injury, spinalmuscular atrophy, spondyloarthropathies, Reiter's syndrome, spontaneousfetal loss, recurrent fetal loss, pre-eclampsia, synucleinopathy,Takayasu's arteritis, post-partum thryoiditis, thyroiditis, Type Icryoglobulinemia, Type II mixed cryoglobulinemia, Type III mixedcryoglobulinemia, ulcerative colitis, uremia, urticaria, venous gasembolus (VGE), or Wegener's granulomatosis; von Hippel-Lindau disease,histoplasmosis of the eye, hard drusen, soft drusen, pigment clumping,or photoreceptor and/or retinal pigmented epithelia (RPE) loss.

Examples of eye disorders that may be treated according to thecompositions and methods disclosed herein include amoebic keratitis,fungal keratitis, bacterial keratitis, viral keratitis, onchorcercalkeratitis, bacterial keratoconjunctivitis, viral keratoconjunctivitis,corneal dystrophic diseases, Fuchs' endothelial dystrophy, Sjogren'ssyndrome, Stevens-Johnson syndrome, autoimmune dry eye diseases,environmental dry eye diseases, corneal neovascularization diseases,post-corneal transplant rejection prophylaxis and treatment, autoimmuneuveitis, infectious uveitis, posterior uveitis (includingtoxoplasmosis), pan-uveitis, an inflammatory disease of the vitreous orretina, endophthalmitis prophylaxis and treatment, macular edema,macular degeneration, age related macular degeneration, proliferativeand non-proliferative diabetic retinopathy, hypertensive retinopathy, anautoimmune disease of the retina, primary and metastatic intraocularmelanoma, other intraocular metastatic tumors, open angle glaucoma,closed angle glaucoma, pigmentary glaucoma and combinations thereof.

In other embodiments, the disorder is selected from glaucoma, diabeticretinopathy, blistering cutaneous diseases (including bullouspemphigoid, pemphigus, and epidermolysis bullosa), ocular cicatricalpemphigoid, uveitis, adult macular degeneration, diabetic retinoparetinitis pigmentosa, macular edema, diabetic macular edema, Behcet'suveitis, multifocal choroiditis, Vogt-Koyangi-Harada syndrome,imtermediate uveitis, birdshot retino-chorioditis, sympatheticophthalmia, ocular dicatricial pemphigoid, ocular pemphigus, nonarterticischemic optic neuropathy, postoperative inflammation, and retinal veinocclusion, or central retinal vein occulusion (CVRO).

Disorders that may be treated or prevented by the degrader or its saltor composition as described herein also include, but are not limited to:hereditary angioedema, capillary leak syndrome, hemolytic uremicsyndrome (HUS), neurological disorders, Guillain Barre Syndrome,diseases of the central nervous system and other neurodegenerativeconditions, glomerulonephritis (including membrane proliferativeglomerulonephritis), SLE nephritis, proliferative nephritis, liverfibrosis, tissue regeneration and neural regeneration, orBarraquer-Simons Syndrome; inflammatory effects of sepsis, systemicinflammatory response syndrome (SIRS), disorders of inappropriate orundesirable complement activation, interleukin-2 induced toxicity duringIL-2 therapy, inflammatory disorders, inflammation of autoimmunediseases, system lupus erythematosus (SLE), lupus nephritides,arthritis, immune complex disorders and autoimmune diseases, systemiclupus, or lupus erythematosus; ischemia/reperfusion injury (I/R injury),myocardial infarction, myocarditis, post-ischemic reperfusionconditions, balloon angioplasty, atherosclerosis, post-pump syndrome incardiopulmonary bypass or renal bypass, renal ischemia, mesentericartery reperfusion after aortic reconstruction, antiphospholipidsyndrome, autoimmune heart disease, ischemia-reperfusion injuries,obesity, or diabetes; Alzheimer's dementia, stroke, schizophrenia,traumatic brain injury, trauma, Parkinson's disease, epilepsy,transplant rejection, prevention of fetal loss, biomaterial reactions(e.g. in hemodialysis, inplants), hyperacute allograft rejection,xenograft rejection, transplantation, psoriasis, burn injury, thermalinjury including burns or frostbite, or crush injury; asthma, allergy,acute respiratory distress syndrome (ARDS), cystic fibrosis, adultrespiratory distress syndrome, dyspnea, hemoptysis, chronic obstructivepulmonary disease (COPD), emphysema, pulmonary embolisms and infarcts,pneumonia, fibrogenic dust diseases, inert dusts and minerals (e.g.,silicon, coal dust, beryllium, and asbestos), pulmonary fibrosis,organic dust diseases, chemical injury (due to irritant gases andchemicals, e.g., chlorine, phosgene, sulfur dioxide, hydrogen sulfide,nitrogen dioxide, ammonia, and hydrochloric acid), smoke injury, thermalinjury (e.g., burn, freeze), bronchoconstriction, hypersensitivitypneumonitis, parasitic diseases, Goodpasture's Syndrome (anti-glomerularbasement membrane nephritis), pulmonary vasculitis, Pauci-immunevasculitis, or immune complex-associated inflammation.

In another embodiment, a method for the treatment of sickle cell in ahost is provided that includes the administration of an effective amountof the degrader or its salt or composition as described herein. In oneembodiment, a method for the treatment of immunothrombocytopenic purpura(ITP), thrombotic thrombocytopenic purpura (TTP), or idiopathicthrombocytopenic purpura (ITP) in a host is provided that includes theadministration of an effective amount of the degrader or its salt orcomposition as described herein. In one embodiment, a method for thetreatment of ANCA-vasculitis in a host is provided that includes theadministration of an effective amount of the degrader or its salt orcomposition as described herein. In one embodiment, a method for thetreatment of IgA nephropathy in a host is provided that includes theadministration of an effective amount of the degrader or its salt orcomposition as described herein. In one embodiment, a method for thetreatment of rapidly progressing glomerulonephritis (RPGN), in a host isprovided that includes the administration of an effective amount of thedegrader or its salt or composition as described herein. In oneembodiment, a method for the treatment of lupus nephritis, in a host isprovided that includes the administration of an effective amount of thedegrader or its salt or composition as described herein. In oneembodiment, a method for the treatment of hemorraghic dengue fever, in ahost is provided that includes the administration of an effective amountof the degrader or its salt or composition as described herein.

In another aspect, an effective amount of the degrader or its salt orcomposition as described herein is used to treat an abnormalproliferation disorder such as a tumor or cancer.

Non-limiting examples of cancers that can be treated according to thepresent invention include, but are not limited to, acoustic neuroma,adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g.,lymphangiosarcoma, lymphangioendotheliosarcoma, hemangiosarcoma),appendix cancer, benign monoclonal gammopathy, biliary cancer (e.g.,cholangiocarcinoma), bladder cancer, breast cancer (e.g., adenocarcinomaof the breast, papillary carcinoma of the breast, mammary cancer,medullary carcinoma of the breast), brain cancer (e.g., meningioma;glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchuscancer, carcinoid tumor, cervical cancer (e.g., cervicaladenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma,colorectal cancer (e.g., colon cancer, rectal cancer, colorectaladenocarcinoma), epithelial carcinoma, ependymoma, endotheliosarcoma(e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma),endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophagealcancer (e.g., adenocarcinoma of the esophagus, Barrett's adenocarinoma),Ewing's sarcoma, eye cancer (e.g., intraocular melanoma,retinoblastoma), familiar hypereosinophilia, gall bladder cancer,gastric cancer (e.g., stomach adenocarcinoma), gastrointestinal stromaltumor (GIST), head and neck cancer (e.g., head and neck squamous cellcarcinoma, oral cancer (e.g., oral squamous cell carcinoma (OSCC),throat cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngealcancer, oropharyngeal cancer)), hematopoietic cancers (e.g., leukemiasuch as acute lymphocytic leukemia (ALL)—also known as acutelymphoblastic leukemia or acute lymphoid leukemia (e.g., B-cell ALL,T-cell ALL), acute myelocytic leukemia (AML) (e.g., B-cell AML, T-cellAML), chronic myelocytic leukemia (CML) (e.g., B-cell CML, T-cell CML),and chronic lymphocytic leukemia (CLL) (e.g., B-cell CLL, T-cell CLL);lymphoma such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) andnon-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large celllymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)),follicular lymphoma, chronic lymphocytic leukemia/small lymphocyticlymphoma (CLL/SLL), mantle cell lymphoma (MCL), marginal zone B-celllymphomas (e.g., mucosa-associated lymphoid tissue (MALT) lymphomas,nodal marginal zone B-cell lymphoma, splenic marginal zone B-celllymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma,lymphoplasmacytic lymphoma (i.e., “Waldenstrom's macroglobulinemia”),hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursorB-lymphoblastic lymphoma and primary central nervous system (CNS)lymphoma; and T-cell NHL such as precursor T-lymphoblasticlymphoma/leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneousT-cell lymphoma (CTCL) (e.g., mycosis fungiodes, Sezary syndrome),angioimmunoblastic T-cell lymphoma, extranodal natural killer T-celllymphoma, enteropathy type T-cell lymphoma, subcutaneouspanniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); amixture of one or more leukemia/lymphoma as described above; andmultiple myeloma (MM)), heavy chain disease (e.g., alpha chain disease,gamma chain disease, mu chain disease), hemangioblastoma, inflammatorymyofibroblastic tumors, immunocytic amyloidosis, kidney cancer (e.g.,nephroblastoma a.k.a. Wilms' tumor, renal cell carcinoma), liver cancer(e.g., hepatocellular cancer (HCC), malignant hepatoma), lung cancer(e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-smallcell lung cancer (NSCLC), adenocarcinoma of the lung), leiomyosarcoma(LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplasticsyndrome (MDS), mesothelioma, myeloproliferative disorder (MPD) (e.g.,polycythemia Vera (PV), essential thrombocytosis (ET), agnogenic myeloidmetaplasia (AMM) a.k.a. myelofibrosis (MF), chronic idiopathicmyelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilicleukemia (CNL), hypereosinophilic syndrome (HES)), neuroblastoma,neurofibroma (e.g., neurofibromatosis (NF) type 1 or type 2,schwannomatosis), neuroendocrine cancer (e.g., gastroenteropancreaticneuroendoctrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovariancancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarianadenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g.,pancreatic andenocarcinoma, intraductal papillary mucinous neoplasm(IPMN), Islet cell tumors), penile cancer (e.g., Paget's disease of thepenis and scrotum), pinealoma, primitive neuroectodermal tumor (PNT),prostate cancer (e.g., prostate adenocarcinoma), rectal cancer,rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamouscell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cellcarcinoma (BCC)), small bowel cancer (e.g., appendix cancer), softtissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma,malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma,fibrosarcoma, myxosarcoma), sebaceous gland carcinoma, sweat glandcarcinoma, synovioma, testicular cancer (e.g., seminoma, testicularembryonal carcinoma), thyroid cancer (e.g., papillary carcinoma of thethyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer),urethral cancer, vaginal cancer and vulvar cancer (e.g., Paget's diseaseof the vulva).

In another embodiment, the disorder is myelodysplastic syndrome (MDS).

In certain embodiments, the cancer is a hematopoietic cancer. In certainembodiments, the hematopoietic cancer is a lymphoma. In certainembodiments, the hematopoietic cancer is a leukemia. In certainembodiments, the leukemia is acute myelocytic leukemia (AML).

In certain embodiments, the proliferative disorder is amyeloproliferative neoplasm. In certain embodiments, themyeloproliferative neoplasm (MPN) is primary myelofibrosis (PMF).

In certain embodiments, the cancer is a solid tumor. A solid tumor, asused herein, refers to an abnormal mass of tissue that usually does notcontain cysts or liquid areas. Different types of solid tumors are namedfor the type of cells that form them. Examples of classes of solidtumors include, but are not limited to, sarcomas, carcinomas, andlymphomas, as described above herein. Additional examples of solidtumors include, but are not limited to, squamous cell carcinoma, coloncancer, breast cancer, prostate cancer, lung cancer, liver cancer,pancreatic cancer, and melanoma.

Abnormal cellular proliferation, notably hyperproliferation, can occuras a result of a wide variety of factors, including genetic mutation,infection, exposure to toxins, autoimmune disorders, and benign ormalignant tumor induction.

There are a number of skin disorders associated with cellularhyperproliferation. Psoriasis, for example, is a benign disease of humanskin generally characterized by plaques covered by thickened scales. Thedisease is caused by increased proliferation of epidermal cells ofunknown cause. Chronic eczema is also associated with significanthyperproliferation of the epidermis. Other diseases caused byhyperproliferation of skin cells include atopic dermatitis, lichenplanus, warts, pemphigus vulgaris, actinic keratosis, basal cellcarcinoma and squamous cell carcinoma.

Other hyperproliferative cell disorders include blood vesselproliferation disorders, fibrotic disorders, autoimmune disorders,graft-versus-host rejection, tumors and cancers.

Blood vessel proliferative disorders include angiogenic and vasculogenicdisorders. Proliferation of smooth muscle cells in the course ofdevelopment of plaques in vascular tissue cause, for example,restenosis, retinopathies and atherosclerosis. Both cell migration andcell proliferation play a role in the formation of atheroscleroticlesions.

Fibrotic disorders are often due to the abnormal formation of anextracellular matrix. Examples of fibrotic disorders include hepaticcirrhosis and mesangial proliferative cell disorders. Hepatic cirrhosisis characterized by the increase in extracellular matrix constituentsresulting in the formation of a hepatic scar. Hepatic cirrhosis cancause diseases such as cirrhosis of the liver. An increasedextracellular matrix resulting in a hepatic scar can also be caused byviral infection such as hepatitis. Lipocytes appear to play a major rolein hepatic cirrhosis.

Mesangial disorders are brought about by abnormal proliferation ofmesangial cells. Mesangial hyperproliferative cell disorders includevarious human renal diseases, such as glomerulonephritis, diabeticnephropathy, malignant nephrosclerosis, thrombotic micro-angiopathysyndromes, transplant rejection, and glomerulopathies.

Another disease with a proliferative component is rheumatoid arthritis.Rheumatoid arthritis is generally considered an autoimmune disease thatis thought to be associated with activity of autoreactive T cells, andto be caused by autoantibodies produced against collagen and IgE.

Other disorders that can include an abnormal cellular proliferativecomponent include Bechet's syndrome, acute respiratory distress syndrome(ARDS), ischemic heart disease, post-dialysis syndrome, leukemia,acquired immune deficiency syndrome, vasculitis, lipid histiocytosis,septic shock and inflammation in general.

In certain embodiments, the condition is associated with an immuneresponse.

Cutaneous contact hypersensitivity and asthma are just two examples ofimmune responses that can be associated with significant morbidity.Others include atopic dermatitis, eczema, Sjogren's Syndrome, includingkeratoconjunctivitis sicca secondary to Sjogren's Syndrome, alopeciaareata, allergic responses due to arthropod bite reactions, Crohn'sdisease, aphthous ulcer, iritis, conjunctivitis, keratoconjunctivitis,ulcerative colitis, cutaneous lupus erythematosus, scleroderma,vaginitis, proctitis, and drug eruptions. These conditions may result inany one or more of the following symptoms or signs: itching, swelling,redness, blisters, crusting, ulceration, pain, scaling, cracking, hairloss, scarring, or oozing of fluid involving the skin, eye, or mucosalmembranes.

In atopic dermatitis, and eczema in general, immunologically mediatedleukocyte infiltration (particularly infiltration of mononuclear cells,lymphocytes, neutrophils, and eosinophils) into the skin importantlycontributes to the pathogenesis of these diseases. Chronic eczema alsois associated with significant hyperproliferation of the epidermis.Immunologically mediated leukocyte infiltration also occurs at sitesother than the skin, such as in the airways in asthma and in the tearproducing gland of the eye in keratoconjunctivitis sicca.

In other non-limiting embodiments, degraders of the present inventionare used as topical agents in treating contact dermatitis, atopicdermatitis, eczematous dermatitis, psoriasis, Sjogren's Syndrome,including keratoconjunctivitis sicca secondary to Sjogren's Syndrome,alopecia areata, allergic responses due to arthropod bite reactions,Crohn's disease, aphthous ulcer, iritis, conjunctivitis,keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma,cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, anddrug eruptions. The novel method may also be useful in reducing theinfiltration of skin by malignant leukocytes in diseases such as mycosisfungoides. These compounds can also be used to treat anaqueous-deficient dry eye state (such as immune mediatedkeratoconjunctivitis) in a patient suffering therefrom, by administeringthe compound topically to the eye.

Exemplary cancers which may be treated by the present disclosedcompounds either alone or in combination with at least one additionalanti-cancer agent include squamous-cell carcinoma, basal cell carcinoma,adenocarcinoma, hepatocellular carcinomas, and renal cell carcinomas,cancer of the bladder, bowel, breast, cervix, colon, esophagus, head,kidney, liver, lung, neck, ovary, pancreas, prostate, and stomach;leukemias; benign and malignant lymphomas, particularly Burkitt'slymphoma and Non-Hodgkin's lymphoma; benign and malignant melanomas;myeloproliferative diseases; sarcomas, including Ewing's sarcoma,hemangiosarcoma, Kaposi's sarcoma, liposarcoma, myosarcomas, peripheralneuroepithelioma, synovial sarcoma, gliomas, astrocytomas,oligodendrogliomas, ependymomas, gliobastomas, neuroblastomas,ganglioneuromas, gangliogliomas, medulloblastomas, pineal cell tumors,meningiomas, meningeal sarcomas, neurofibromas, and Schwannomas; bowelcancer, breast cancer, prostate cancer, cervical cancer, uterine cancer,lung cancer, ovarian cancer, testicular cancer, thyroid cancer,astrocytoma, esophageal cancer, pancreatic cancer, stomach cancer, livercancer, colon cancer, melanoma; carcinosarcoma, Hodgkin's disease,Wilms' tumor and teratocarcinomas. Additional cancers which may betreated using the disclosed compounds according to the present inventioninclude, for example, acute granulocytic leukemia, acute lymphocyticleukemia (ALL), acute myelogenous leukemia (AML), adenocarcinoma,adenosarcoma, adrenal cancer, adrenocortical carcinoma, anal cancer,anaplastic astrocytoma, angiosarcoma, appendix cancer, astrocytoma,Basal cell carcinoma, B-Cell lymphoma, bile duct cancer, bladder cancer,bone cancer, bone marrow cancer, bowel cancer, brain cancer, brain stemglioma, breast cancer, triple (estrogen, progesterone and HER-2)negative breast cancer, double negative breast cancer (two of estrogen,progesterone and HER-2 are negative), single negative (one of estrogen,progesterone and HER-2 is negative), estrogen-receptor positive,HER2-negative breast cancer, estrogen receptor-negative breast cancer,estrogen receptor positive breast cancer, metastatic breast cancer,luminal A breast cancer, luminal B breast cancer, Her2-negative breastcancer, HER2-positive or negative breast cancer, progesteronereceptor-negative breast cancer, progesterone receptor-positive breastcancer, recurrent breast cancer, carcinoid tumors, cervical cancer,cholangiocarcinoma, chondrosarcoma, chronic lymphocytic leukemia (CLL),chronic myelogenous leukemia (CML), colon cancer, colorectal cancer,craniopharyngioma, cutaneous lymphoma, cutaneous melanoma, diffuseastrocytoma, ductal carcinoma in situ (DCIS), endometrial cancer,ependymoma, epithelioid sarcoma, esophageal cancer, ewing sarcoma,extrahepatic bile duct cancer, eye cancer, fallopian tube cancer,fibrosarcoma, gallbladder cancer, gastric cancer, gastrointestinalcancer, gastrointestinal carcinoid cancer, gastrointestinal stromaltumors (GIST), germ cell tumor glioblastoma multiforme (GBM), glioma,hairy cell leukemia, head and neck cancer, hemangioendothelioma, Hodgkinlymphoma, hypopharyngeal cancer, infiltrating ductal carcinoma (IDC),infiltrating lobular carcinoma (ILC), inflammatory breast cancer (IBC),intestinal Cancer, intrahepatic bile duct cancer, invasive/infiltratingbreast cancer, Islet cell cancer, jaw cancer, Kaposi sarcoma, kidneycancer, laryngeal cancer, leiomyosarcoma, leptomeningeal metastases,leukemia, lip cancer, liposarcoma, liver cancer, lobular carcinoma insitu, low-grade astrocytoma, lung cancer, lymph node cancer, lymphoma,male breast cancer, medullary carcinoma, medulloblastoma, melanoma,meningioma, Merkel cell carcinoma, mesenchymal chondrosarcoma,mesenchymous, mesothelioma metastatic breast cancer, metastatic melanomametastatic squamous neck cancer, mixed gliomas, monodermal teratoma,mouth cancer mucinous carcinoma, mucosal melanoma, multiple myeloma,Mycosis Fungoides, myelodysplastic syndrome, nasal cavity cancer,nasopharyngeal cancer, neck cancer, neuroblastoma, neuroendocrine tumors(NETs), non-Hodgkin's lymphoma, non-small cell lung cancer (NSCLC), oatcell cancer, ocular cancer, ocular melanoma, oligodendroglioma, oralcancer, oral cavity cancer, oropharyngeal cancer, osteogenic sarcoma,osteosarcoma, ovarian cancer, ovarian epithelial cancer ovarian germcell tumor, ovarian primary peritoneal carcinoma, ovarian sex cordstromal tumor, Paget's disease, pancreatic cancer, papillary carcinoma,paranasal sinus cancer, parathyroid cancer, pelvic cancer, penilecancer, peripheral nerve cancer, peritoneal cancer, pharyngeal cancer,pheochromocytoma, pilocytic astrocytoma, pineal region tumor,pineoblastoma, pituitary gland cancer, primary central nervous system(CNS) lymphoma, prostate cancer, rectal cancer, renal cell carcinoma,renal pelvis cancer, rhabdomyosarcoma, salivary gland cancer, softtissue sarcoma, bone sarcoma, sarcoma, sinus cancer, skin cancer, smallcell lung cancer (SCLC), small intestine cancer, spinal cancer, spinalcolumn cancer, spinal cord cancer, squamous cell carcinoma, stomachcancer, synovial sarcoma, T-cell lymphoma, testicular cancer, throatcancer, thymoma/thymic carcinoma, thyroid cancer, tongue cancer, tonsilcancer, transitional cell cancer, tubal cancer, tubular carcinoma,undiagnosed cancer, ureteral cancer, urethral cancer, uterineadenocarcinoma, uterine cancer, uterine sarcoma, vaginal cancer, vulvarcancer, T-cell lineage acute lymphoblastic leukemia (T-ALL), T-celllineage lymphoblastic lymphoma (T-LL), peripheral T-cell lymphoma, AdultT-cell leukemia, Pre-B ALL, Pre-B lymphomas, large B-cell lymphoma,Burkitts lymphoma, B-cell ALL, Philadelphia chromosome positive ALL,Philadelphia chromosome positive CML, juvenile myelomonocytic leukemia(JMML), acute promyelocytic leukemia (α subtype of AML), large granularlymphocytic leukemia, Adult T-cell chronic leukemia, diffuse large Bcell lymphoma, follicular lymphoma; Mucosa-Associated Lymphatic Tissuelymphoma (MALT), small cell lymphocytic lymphoma, mediastinal large Bcell lymphoma, nodal marginal zone B cell lymphoma (NMZL); splenicmarginal zone lymphoma (SMZL); intravascular large B-cell lymphoma;primary effusion lymphoma; or lymphomatoid granulomatosis; B-cellprolymphocytic leukemia; splenic lymphoma/leukemia, unclassifiable,splenic diffuse red pulp small B-cell lymphoma; lymphoplasmacyticlymphoma; heavy chain diseases, for example, Alpha heavy chain disease,Gamma heavy chain disease, Mu heavy chain disease, plasma cell myeloma,solitary plasmacytoma of bone; extraosseous plasmacytoma; primarycutaneous follicle center lymphoma, T cell/histocyte rich large B-celllymphoma, DLBCL associated with chronic inflammation; Epstein-Barr virus(EBV)+DLBCL of the elderly; primary mediastinal (thymic) large B-celllymphoma, primary cutaneous DLBCL, leg type, ALK+large B-cell lymphoma,plasmablastic lymphoma; large B-cell lymphoma arising in HHV8-associatedmulticentric, Castleman disease; B-cell lymphoma, unclassifiable, withfeatures intermediate between diffuse large B-cell lymphoma, or B-celllymphoma, unclassifiable, with features intermediate between diffuselarge B-cell lymphoma and classical Hodgkin lymphoma.

Nonlimiting general examples of disorders mediated by extracellularproteins also include, but are not limited to: AMD, macular edema, DME,diabetic retinopathy, mCNV; neurodegenerative disorders, metastaticcolorectal cancer, non-squamous non-small-cell lung carcinoma, GMB,metastatic renal cell carcinoma, cervical cancer, AA amyloidosis,amyloid light chain (AL) amyloidosis, ankylosing spondylitis,antiphospholipid Ab syndrome, asthma, progression of parasiteschistosoma Mansoni infection (IL-13), ATTR amyloidosis, Behcetsyndrome, sepsis, inflammation, rheumatoid arthritis, atherosclerosis,ischemia/reperfusion injury; MGUS, Necrobiotic xanthogranuloma, JIA,psoriatic arthritis, plaque psoriasis, Crohn's disease, ulcerativecolitis, Hidradenitis suppurativa uveitis; GvH disease; Castleman'sdisease, liver fibrosis, Still's Disease; cutaneous skin diseasesincluding atopic dermatitis, transplant rejection, multiple myeloma,osteosclerotic multiple myeloma with peripheral neuropathy; pancreatictumors; paraproteinemia (NR), prostate, gastric cancer; glioblastomamultiforme; acute coronary syndrome; hyperlipidemia (Rare/Broad),chronic urticaria, scleroderma, scleromyxedema, hereditary angioedema,clotting disorders, heparin-induced thrombocytopenia; Acquired VonWillebrand disease (AVWD), antiphospholipid antibody syndrome (APS orAPLS); cryoglobulinemia; granulomatosis with polyangiitis(Wegener's)—sub-type of ANCA-associated vasculitis; idiopathic (Immune);thrombocytopenic purpura; IgG4-RD; Non-IgM MGUS; X-linkedhypophosphatemia; Multiple System Atrophy (MSA), Parkinson's disease,Cachexia, Sarcopenia, Sporadic inclusion body myositis, musculardystrophy, COPD; rhabdomyolysis; dialysis-related amyloidosis; focalsegmental glomerulosclerosis (FSGS); IgA nephropathy (IgAN) and HenochSchonlein Purpura (HSP); acute disseminated encephalomyelitis (ADEM);acute inflammatory demyelinating polyneuropathy (AIDP); Guillaine-BarreSyndrome; Alzheimer'disease & FTD; chronic inflammatory demyelinatingpolyneuropathy (CIDP); Creutzfeldt-Jakob disease (CJD); Huntington'sdisease; Miller Fisher Syndrome; Neuromyelitis optica spectrum disorder(NMOSD); Opsoclonus-myoclonus syndrome; PANDAS syndrome (pediatricautoimmune neuropsychiatric disorders associated with Streptcoccalinfections); Transverse myelitis; Emphysema, respiratory failure;Anthrax; Botulism; Sepsis; Staph. aureus toxic shock syndrome; Tetanus;Transplantation; Acromegaly; Cushing's disease; prion disease; secondarymembranous nephropathy; and vasculitis.

IX. Processes of Manufacture:

The extracellular protein degrading compounds of the present inventioncan be manufactured according to routes described in the WorkingExamples below or as otherwise known in the patent or scientificliterature and if appropriate supported by the knowledge of the ordinaryworker or common general knowledge.

Some of the carbons in the extracellular protein degrading compoundsdescribed herein are drawn with designated stereochemistry. Othercarbons are drawn without stereochemical designation. When drawn withoutdesignated stereochemistry, that carbon can be in any desiredstereochemical configuration that achieves the desired purpose. Oneskilled in the art will recognize that pure enantiomers,enantiomerically enriched compounds, racemates and diastereomers can beprepared by methods known in the art as guided by the informationprovided herein. Examples of methods to obtain optically activematerials include at least the following:

-   -   i) chiral liquid chromatography—a technique whereby        diastereomers are separated in a liquid mobile phase by virtue        of their differing interactions with a stationary phase        (including vial chiral HPLC). The stationary phase can be made        of chiral material or the mobile phase can contain an additional        chiral material to provoke the differing interactions;    -   ii) non-chiral chromatography of diastereomers-Often        diastereomers can be separated using normal non-chiral column        conditions;    -   iii) chiral gas chromatography—a technique whereby the racemate        is volatilized and enantiomers are separated by virtue of their        differing interactions in the gaseous mobile phase with a column        containing a fixed non-racemic chiral adsorbent phase;    -   iv) simultaneous crystallization—a technique whereby the        individual diastereomers are separately crystallized from a        solution;    -   v) enzymatic resolutions—a technique whereby partial or complete        separation of diastereomers are separated by virtue of differing        rates of reaction with an enzyme;    -   vi) chemical asymmetric synthesis—a synthetic technique whereby        the desired diastereomer is synthesized from an achiral        precursor under conditions that produce asymmetry (i.e.        chirality) in the product, which may be achieved by chiral        catalysts or chiral auxiliaries;    -   vii) diastereomer separations—a technique whereby a racemic        compound is reaction with an enantiomerically pure reagent (the        chiral auxiliary) that converts the individual enantiomers to        diastereomers. The resulting diastereomers are then separated by        chromatography or crystallization by virtue of their now more        distinct structural differences the chiral auxiliary later        removed to obtain the desired enantiomer; and    -   viii) extraction with chiral solvents—a technique whereby        diastereomers are separated by virtue of preferential        dissolution of one over the others in a particular chiral        solvent.        General Procedures applied to the working examples of synthesis:        All reagents were purchased from commercial suppliers        (Sigma-Aldrich, Alfa, Across etc.) and used without further        purification unless otherwise stated. THE was continuously        refluxed and freshly distilled from sodium and benzophenone        under nitrogen, dichloromethane was continuously refluxed and        freshly distilled from CaH₂ under nitrogen.        Reactions were monitored via TLC on silica gel 60 HSGF254        percolated plates (0.15-0.2 mm SiO₂) and visualized using UV        light (254 nm or 365 nm) and/or staining with phosphomolybdic        acid ethanol solution (10 g in 100 mL ethanol) and subsequent        heating or monitored via LCMS.

LCMS were performed on SHIMADZU LCMS—2010EV (Chromolith SpeedROD,RP-18e, 50×4.6 mm, mobile phase: Solvent A: CH₃CN/H₂O/HCOOH=10/90/0.05,Solvent B: CH₃CN/H₂O/HCOOH=90/10/0.05, 0.8 min@ 10% B, 2.7 min gradient(10-95% B), then 0.8 min@95% B, Flow rate: 3 mL/min, temperature: 40°C.).

Preparative HPLC were performed either on Method A: SHIMADZU LC-8A(Column: YMC Pack ODS-A (150*30 mm, 10 m)) or Method B: LC-6AD (Column:Shim=Pack PREP-ODS-H (250*20 mm, 10 m)) with UV detection which werecontrolled by LC solution Chemstation software. H₂O (0.1% HCOOH) andMeOH (MeCN) as mobile phase at the indicated flow rate.Analytical HPLC were performed on SHIMADZU LC-2010A (ChromolithSpeedROD, RP-18e, 50×4.6 mm, mobile phase: Solvent A:CH₃CN/H₂O/HCOOH=10/90/0.05, Solvent B: CH₃CN/H₂O/HCOOH=90/10/0.05, 0.8min@ 10% B, 2.7 min gradient (10-95% B), then 0.8 min@95% B, Flow rate:3 mL/min, temperature: 40° C.).Chiral HPLC were performed on SHIMADZU LC-2010A (Chiral column, mobilephase: Solvent A: hexane (or contained 0.1% diethylamine), Solvent B:Ethanol or Isopropanol; Flow rate: 0.8 mL/min, temperature: 30° C.).¹H spectra were recorded on Bruker Avance II 400 MHz, Chemical shifts(6) were reported in ppm relative to tetramethylsilane (S=0.000 ppm) andthe spectra were calibrated to the residual solvent signal of chloroform(S=7.26),Dimethyl sulfoxide (S=2.50), methanol (S=3.30). Data for ¹H NMRspectra were reported as following: chemical shift (multiplicity, numberof hydrogens).

Abbreviations were described as following: s (singlet), d (doublet), t(triplet), q (quartet), quant (quintet), m (multiple), br (broad).

X. WORKING EXAMPLES Example 1. Synthesis of Boc-Protected andBn-Protected Intermediates Synthesis 1-1. Preparation of tert-Butyl(((3aR,4S,8R,8aR)-8-amino-2,2-dimethyltetrahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4(5H)-yl)methyl)carbamate(Intermediate 1)

Synthesis 1-2. Preparation of tert-Butyl(((3aS,4R,7R,7aR)-7-amino-6-hydroxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)carbamate(Intermediate 2)

Synthesis 1-3: Preparation of(2R,3R,4R,5R,6R)-4,5-Bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(Intermediate 3)

Step 1: To a stirred mixture of 1 mL methanol (24.5 mmol) and TEA (5 mL)was added a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-nitro-3,4-dihydro-2H-pyran(1.0 g, 2.17 mmol) in dry THF (5 mL) dropwise under argon. Afterstirring for 5 h at rt, the volatiles were removed in vacuo. Theresulting crude material was purified by column to give(2R,3R,4R,5R,6R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-6-methoxy-5-nitrotetrahydro-2H-pyran(535 mg, 50%) and(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-nitro-3,4-dihydro-2H-pyran(65 mg, 6%). LC-MS (ESI) of both found: 494 [M+H]⁺.

Step 2: To a solution of(2R,3R,4R,5R,6R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-6-methoxy-5-nitrotetrahydro-2H-pyran(535 mg, 1.09 mmol) in MeOH (10 mL) was added Raney Ni (50 mg). Themixture was charged with H₂ for three times and stirred at rt for 12 hunder a H₂ balloon. The mixture was filtered and the filtrate wasconcentrated to give(2R,3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(300 mg, 59% yield) as white solid. LC-MS (ESI) found: 464 [M+H]⁺.

Synthesis 1-4: Preparation ofN-((3aR,4R,6R,7R,7aR)-4-(Hydroxymethyl)-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(Intermediate 4)

Step 1: To a solution of(2R,3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(1.5 g, 3.24 mmol) in DCM (15 mL) was added AcCl (508 mg, 6.48 mmol) andTEA (3 mL). The mixture was stirred at rt for 2 h. The mixture wasconcentrated and purified by silica column to giveN-((2R,3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(1.6 g, 98%) as white solid. LC-MS (ESI) found: 506 [M+H]⁺.

Step 2: To a solution ofN-((2R,3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(1.6 g, 3.17 mmol) in MeOH (15 mL) was added Pd/C (100 mg, 10% wt, 60%wet). The mixture was stirred at rt for 12 h under a H₂ balloon. Themixture was filtered and concentrated to giveN-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(670 mg, 90% yield). LC-MS (ESI) found: 236 [M+H]⁺. ¹H NMR (400 MHz,CD₃OD): δ4.68 (d, J=3.7 Hz, 1H), 4.27 (dd, J=10.9, 3.6 Hz, 1H), 3.87 (d,J=3.1 Hz, 1H), 3.79-3.69 (m, 3H), 3.37 (d, J=5.6 Hz, 3H), 2.02-1.93 (m,3H).

Step 3: To a solution ofN-((2S,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(670 mg, 2.85 mmol) in DMF (5 mL) and 2,2-dimethoxypropane (0.8 mL, 6.42mmol) were added (+/−)-camphor-10-sulphonic acid (330 mg, 1.42 mmol).The reaction mixture was stirred at 70° C. for 24 h. Then it was cooledto room temperature and neutralized with triethylamine. The solvent wasevaporated and the residue was co-evaporated 3 times with toluene. Theresulting crude material was purified by column to giveN-((3aR,4R,6S,7R,7aR)-4-(hydroxymethyl)-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(392 mg, 50%) as white solid. LC-MS (ESI) found: 276 [M+H]⁺. ¹H NMR (400MHz, CD3OD): δ4.30 (dd, J=8.6, 5.5 Hz, 1H), 4.16 (ddd, J=13.6, 6.8, 3.5Hz, 2H), 3.88-3.71 (m, 3H), 3.48-3.41 (m, 3H), 3.34 (s, 1H), 2.01-1.90(m, 3H), 1.53-1.44 (m, 3H), 1.32 (d, J=11.4 Hz, 3H).

Example 2. Synthesis of ASGPR Ligands Synthesis 2-1. General Synthesisof Sulfonamide-containing Ligands

Synthesis 2-2. General Synthesis of Sulfonimidamide-containing Ligands

Synthesis 2-3. General Synthesis of Sulfonyl Urea-Containing Compounds

Synthesis 2-4: Alternative General Synthesis of Sulfonyl Urea-ContainingCompounds

Synthesis 2-5. General Synthesis of Sulfonimidamide-containing Ligands

Synthesis 2-6. General Synthesis of ASGPR Ligands

Synthesis 2-7. General Synthesis of ASGPR Ligands

Synthesis 2-6 and Synthesis 2-7 can be used to synthesize ligands withthe following R² groups:

wherein R is an optimal substituent has defined herein.Synthesis 2-8. Preparation ofN-(((3R,4R,5R,6R)-2,4,5-Trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(Compound A9) andN-(((3R,4R,5R,6R)-6-(aminomethyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-yl)methyl)acetamide(Compound A10)

Synthesis 2-9. General Synthesis of Amide-Containing Ligands

Synthesis 2-10. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)pyrrolidin-2-one(Compound A15)

Synthesis 2-11. Preparation of3-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)oxazolidin-2-one(Compound A16)

Synthesis 2-12. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)imidazolidin-2-one(Compound A17)

Synthesis 2-13. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)imidazolidine-2-thione(Compound A18)

Synthesis 2-14. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)pyrrolidine-2,5-dione(Compound A19)

Synthesis 2-15. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)-1H-pyrrole-2,5-dione(Compound A20)

Synthesis 2-16. Preparation of3-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)thiazolidine-2,4-dione(Compound A21)

Synthesis 2-17. Preparation of3-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)oxazolidine-2,4-dione(Compound A22)

Synthesis 2-18. Preparation of2-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)isoindoline-1,3-dione(Compound A23)

Synthesis 2-19. Preparation of2-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)isoindolin-1-one(Compound A24)

Synthesis 2-20. Preparation of(1S,2R,3R,4R)-1-(Aminomethyl)-4-(1H-imidazol-1-yl)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(Compound A25)

Synthesis 2-21. Preparation of(1S,2R,3R,4R)-1-(Aminomethyl)-4-(1H-pyrrol-1-yl)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(Compound A26)

Synthesis 2-22. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)pyridin-2(1H)-one(Compound A27)

Synthesis 2-23. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)pyrimidin-2(1H)-one(Compound A28)

Synthesis 2-24. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)pyridin-4(1H)-one(Compound A29)

Synthesis 2-25. Preparation of(3R,4R,5R,6R)-6-(Aminomethyl)-3-(piperidin-1-yl)tetrahydro-2H-pyran-2,4,5-triol(Compound A30)

Synthesis 2-26. Preparation of(3R,4R,5R,6R)-6-(Aminomethyl)-3-morpholinotetrahydro-2H-pyran-2,4,5-triol(Compound A31)

Synthesis 2-27. Preparation of(3R,4R,5R,6R)-6-(Aminomethyl)-3-thiomorpholinotetrahydro-2H-pyran-2,4,5-triol(Compound A32)

Synthesis 2-28. Preparation of1-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)azetidin-2-one(Compound A33)

Synthesis 2-29. Preparation of(4R,5R,6R)-6-(Aminomethyl)tetrahydro-2H-pyran-2,4,5-triol (Compound A34)and (4R,5R,6R)-6-(hydroxymethyl)tetrahydro-2H-pyran-2,4,5-triol(Compound A35)

Synthesis 2-30. Preparation of1-((3R,4R,5R,6R)-6-(Aminomethyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-yl)tetrahydropyrimidin-2(1H)-one(Compound A36)

Synthesis 2-31. Preparation ofN-((1S,2R,3R,4R)-1-(Aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)methanesulfinamide(Compound A37) andN-((1S,2R,3R,4R)-1-(aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)-1,1,1-trifluoromethanesulfinamide(Compound A38)

Synthesis 2-32. Preparation of(4aR,6R,7R,8R,8aR)-6-(Aminomethyl)-7,8-dihydroxyhexahydro-1H,3H-pyrano[3,2-c][1,2,6]thiadiazine2,2-dioxide (Compound A39),(4aR,6R,7R,8R,8aR)-6-(aminomethyl)-7,8-dihydroxyhexahydro-1H-pyrano[3,2-d]pyrimidin-2(3H)-one(Compound A40), and(4aS,6R,7R,8R,8aR)-6-(aminomethyl)-7,8-dihydroxyhexahydropyrano[3,2-d][1,3]oxazin-2(1H)-one(Compound A41)

Synthesis 2-33. Alternative Preparation of(4aS,6R,7R,8R,8aR)-6-(Aminomethyl)-7,8-dihydroxytetrahydro-1H,6H-pyrano[2,3-b][1,4]oxazin-2(3H)-one(Compound A41)

Synthesis 2-34. Preparation of(3aR,5R,6R,7R,7aR)-7-Amino-5-(hydroxymethyl)-2-methyl-3a,6,7,7a-tetrahydro-5H-pyrano[3,2-d]oxazol-6-ol(Compound A43) and(3aR,5R,6R,7R,7aR)-7-amino-5-(hydroxymethyl)-2-(trifluoromethyl)-3a,6,7,7a-tetrahydro-5H-pyrano[3,2-d]oxazol-6-ol(Compound A44)

Synthesis 2-35. Preparation of(5R,6R,7R)-5-(Hydroxymethyl)-2-methyl-6,7-dihydro-5H-pyrano[3,2-d]oxazole-6,7-diol(Compound A45) and(5R,6R,7R)-5-(hydroxymethyl)-2-(trifluoromethyl)-6,7-dihydro-5H-pyrano[3,2-d]oxazole-6,7-diol(Compound A46)

Synthesis 2-36. Preparation of(3aS,5R,6R,7R,7aR)-7-Amino-5-(hydroxymethyl)-2-methyl-3,3a,5,6,7,7a-hexahydropyrano[2,3-d]imidazol-6-ol(Compound A47) and(3aS,5R,6R,7R,7aR)-7-amino-5-(hydroxymethyl)-2-(trifluoromethyl)-3,3a,5,6,7,7α-hexahydropyrano[2,3-d]imidazol-6-ol(Compound A48)

Synthesis 2-37. Preparation of(5R,6R,7R)-5-(Hydroxymethyl)-2-methyl-3,5,6,7-tetrahydropyrano[2,3-d]imidazole-6,7-diol(Compound A49) and(5R,6R,7R)-5-(hydroxymethyl)-2-(trifluoromethyl)-3,5,6,7-tetrahydropyrano[2,3-d]imidazole-6,7-diol(Compound A50)

Synthesis 2-38. Preparation of1-((3aR,5R,6R,7R,7aR)-5-(Aminomethyl)-6,7-dihydroxyhexahydropyrano[3,2-b]pyrrol-1(2H)-yl)-2,2,2-trifluoroethan-1-one(Compound A51) and1-((5R,6R,7R)-5-(aminomethyl)-6,7-dihydroxy-6,7-dihydropyrano[3,2-b]pyrrol-1(5H)-yl)-2,2,2-trifluoroethan-1-one(Compound A52)

Synthesis 2-39. Preparation of1-((4aS,6R,7R,8R,8aR)-6-(Aminomethyl)-7,8-dihydroxyhexahydro-1H,6H-pyrano[2,3-b][1,4]oxazin-1-yl)ethan-1-one(Compound A53) and1-((4aS,6R,7R,8R,8aR)-6-(aminomethyl)-7,8-dihydroxyhexahydro-1H,6H-pyrano[2,3-b][1,4]oxazin-1-yl)-2,2,2-trifluoroethan-1-one(Compound A54)

Synthesis 2-40. Preparation of1-((3aS,4R,5aS,9aR,9bR)-4-(Aminomethyl)-2,2,7-trimethylhexahydro-4H,9H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazin-9-yl)ethan-1-one(Compound A55) and1-((3aS,4R,5aS,9aR,9bR)-4-(Aminomethyl)-2,2,7-trimethylhexahydro-4H,9H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazin-9-yl)-2,2,2-trifluoroethan-1-one(Compound A56)

Synthesis 2-41. Preparation of1-((3aS,4R,5aS,11aR,11bR)-4-(Aminomethyl)-2,2-dimethyloctahydro-4H,11H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazocin-11-yl)ethan-1-one(Compound A57) and1-((3aS,4R,5aS,11aR,11bR)-4-(Aminomethyl)-2,2-dimethyloctahydro-4H,11H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazocin-11-yl)-2,2,2-trifluoroethan-1-one(Compound A58)

Synthesis 2-42. Preparation of1-((3aR,5R,6R,7R,7aR)-6,7-Dihydroxy-5-(hydroxymethyl)hexahydropyrano[3,2-b]pyrrol-1(2H)-yl)ethan-1-one(Compound A59) and(2R,3R,4R,4aR,8aR)-2-(Hydroxymethyl)octahydro-2H-pyrano[3,2-b]pyridine-3,4-diol(Compound A60)

Synthesis 2-43. Preparation of1-((2R,3R,4R,4aR,9aR)-3,4-Dihydroxy-2-(hydroxymethyl)octahydropyrano[3,2-b]azepin-5(2H)-yl)ethan-1-one(Compound A61)

Synthesis 2-44. Preparation of((3aR,4R,5aR,9aS,9bR)-2,2-Dimethyl-8-oxooctahydro-4H-[1,3]dioxolo[4′,5′:4,5]pyrano[3,2-b]pyridin-4-yl)methylacetate (Compound A62)

Synthesis 2-45. Preparation of(2R,3R,4R)-2-(Hydroxymethyl)-3,4-dihydro-2H-pyrano[3,2-b]pyridine-3,4-diol(Compound A63) and((3aR,4R,9bR)-2,2-dimethyl-8-oxooctahydro-4H-[1,3]dioxolo[4′,5′:4,5]pyrano[3,2-b]pyridin-4-yl)methylacetate (Compound A64)

Synthesis 2-46. Preparation ofN-((3aR,8R,8aR)-4-(Hydroxymethyl)-2,2-dimethylhexahydro-4H-4,7-epoxy[1,3]dioxolo[4,5-d]azepin-8-yl)acetamide(Compound A65)

Synthesis 2-47. Preparation ofN-((3aR,4R,9R,9aR)-9-(Hydroxymethyl)-2,2-dimethyloctahydro-5,9-epoxy[1,3]dioxolo[4,5-d]azocin-4-yl)acetamide(Compound A66)

Synthesis 2-48. Preparation ofN-((3aR,4R,9R,9aR)-9-(Hydroxymethyl)-2,2-dimethylhexahydro-5H-5,9-epoxy[1,3]dioxolo[4,5-d]oxocin-4-yl)acetamide(Compound A67)

Compound A68, Compound A69, and Compound A70 can also be synthesizedusing Synthesis 2-46, 2-47, and 2-48.

Synthesis 2-49. Preparation of(3R,4R,5R,6R)-6-(Aminomethyl)-3-(trifluoromethyl)tetrahydro-2H-pyran-2,4,5-triol(Compound A71)

Synthesis 2-50. Preparation of General Synthesis to install R²

Synthesis 2-51. Alternative General Synthesis to install R²

Synthesis 2-52. Preparation of(1S,2R,3R,4R)-1-(Aminomethyl)-4-(isoxazol-5-ylamino)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(Compound A73)

Synthesis 2-53. Preparation of(1S,2R,3R,4R)-1-(Aminomethyl)-4-((4,6-dichloro-1,3,5-triazin-2-yl)amino)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(Compound A74)

Synthesis 2-54. Preparation of(3R,4R,5R,6R)-6-(Aminomethyl)-3-(thiazol-2-ylamino)tetrahydro-2H-pyran-2,4,5-triol(Compound A75)

Synthesis 2-55. Preparation of(2R,3R,4R,5S)-2-(Hydroxymethyl)-5-((3-(trifluoromethyl)pyridin-2-yl)amino)tetrahydro-_2H-pyran-34-diol(Compound A76)

Synthesis 2-56. Preparation of(3aR,4R,8S,8aR)-8-Azido-4-(azidomethyl)-2,2-dimethylhexahydro-4H-4,7-epoxycyclohepta[d][1,3]dioxole(Compound A78)

Synthesis 2-57. Preparation of(3R,4S,5R,6R)-6-(aminomethyl)-3-(oxazol-2-yloxy)tetrahydro-2H-pyran-2,4,5-triol(Compound A79)

Synthesis 2-58. Preparation of(3R,4S,5R,6R)-6-(aminomethyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-ylacetate (Compound A80)

Synthesis 2-59. Preparation of1-((3R,4R,5R,6R)-6-(aminomethyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-yl)guanidine(Compound A81) and (Compound A82)

Synthesis 2-60. Preparation of Compound A83 and Compound A84

Alternatively, Compound A85 can be synthesized if

is used instead of

in the Schotten Bauman reaction step.

Synthesis 2-61. Preparation of Compound A88

Synthesis 2-62: Preparation of(2R,3R,4R,5R,6R)-5-azido-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A89) and(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A90)

Step 1: NaN₃ (4.3 g, 66 mmol) and CAN (87 g, 158 mmol) were added to anitrogen-flushed flask, and the mixture were stirred vigorously at −10°C. Then a solution of(2R,3R,4R)-2-(acetoxymethyl)-3,4-dihydro-2H-pyran-3,4-diyl diacetate(A90-1, 12 g, 44 mmol) in MeCN (250 mL) were added dropwise to the abovemixture. The mixture was stirred at rt for 12 h. Then the reactionmixture was diluted with 500 mL EA. The organic phase was washed withH₂O (400 mL×3) and brine, filtered and concentrated to give a yellowoil, which was purified by column chromatography to give(2R,3R,4R,5R)-2-(acetoxymethyl)-5-azido-6-(nitrooxy)tetrahydro-2H-pyran-3,4-diyldiacetate (A90-2, 7.5 g, 45% yield) as white solid. LC-MS (ESI) found:377 [M+H]⁺.

Step 2: To a solution of(2R,3R,4R,5R)-2-(acetoxymethyl)-5-azido-6-(nitrooxy)tetrahydro-2H-pyran-3,4-diyldiacetate (A90-2, 15.0 g, 40.0 mmol) in anhydrous MeCN (120 mL) wasadded LiBr (34.6 g, 400 mmol) at room temperature under an argonatmosphere.

The reaction was stirred at room temperature for 3 h. TLC indicated thestarting material was consumed. EA (350 mL) was added to the reactionmixture. The organic phase was washed with water (50 mL×2), saturatedNaHCO₃ (60 mL×2), water (50 mL×2), brine (50 mL), dried (Na₂SO₄),filtered. The filtrate was concentrated to give a crude(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-5-azido-6-bromotetrahydro-2H-pyran-3,4-diyldiacetate (A90-3, 15.0 g, 96%) as white foam. LC-MS (ESI) found: 394[M+H]⁺.

Step 3: To a solution of(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-5-azido-6-bromotetrahydro-2H-pyran-3,4-diyldiacetate (A90-3, 15.0 g, 38.1 mmol) in MeOH (100 mL) was added Ag₂CO₃(15.7 g, 57.1 mmol) in portions at rt. The mixture was stirred at 60° C.under N₂ in the dark for 12 h. EA (350 mL) was added to the reactionmixture. The organic phase was washed with water (50 mL×2), saturatedNaHCO₃ (60 mL×2), water (50 mL×2), brine (50 mL), dried (Na₂SO₄),filtered. The filtrate was concentrated to give a crude product, whichwas purified by column chromatography to give(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-5-azido-6-methoxytetrahydro-2H-pyran-3,4-diyldiacetate (A90-4, 10.0 g, 76%) as colorless oil. LC-MS (ESI) found: 346[M+H]⁺. ¹H NMR (400 MHz, CD3OD): δ5.33-5.29 (m, 1H), 4.91-4.86 (m, 1H),4.43 (d, J=8.0 Hz, 1H), 4.16-4.10 (m, 2H), 4.07-4.00 (m, 1H), 3.63-3.59(m, 1H), 3.57 (d, J=3.7 Hz, 3H), 2.15-2.13 (m, 3H), 2.03-1.97 (m, 6H).

Step 4: To a solution of(2R,3R,4R,5R,6R)-2-(acetoxymethyl)-5-azido-6-methoxytetrahydro-2H-pyran-3,4-diyldiacetate (A90-4, 10.0 g, 29.0 mmol) in MeOH (150 mL) was added NaOMe(23.2 mL, 5 M in MeOH) in portions at rt. The mixture was stirred at rtfor 2 h. The reaction was neutralized by the addition of acidicAmberlite IR 120 (H⁺) ion exchange resin. The solution was filteredthrough a glass fritted vacuum filter funnel equipped with a pad ofCelite to remove the acidic resin. The filtrate was concentrated andpurified by column to give(2R,3R,4R,5R,6R)-5-azido-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A89, 5.5 g, 78%) as white solid. LC-MS (ESI) found: 220 [M+H]⁺. ¹H NMR(400 MHz, CD3OD): δ4.21-4.15 (m, 1H), 3.79 (t, J=3.5 Hz, 1H), 3.76-3.70(m, 2H), 3.54 (s, 3H), 3.49-3.42 (m, 1H), 3.41 (dd, J=4.0, 2.6 Hz, 2H).

Step 5: To a solution of(2R,3R,4R,5R,6R)-5-azido-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A-89, 1.0 g, 4.57 mmol) in MeOH (20 mL) was added Pd/C (100 mg, 10% wt,60% wet) under H₂ atmosphere. The mixture was stirred at rt for 12 hunder a H₂ balloon. The mixture was filtered and concentrated to give(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 749 mg, 85% yield) as white solid. LC-MS (ESI) found: 194 [M+H]⁺.

Synthesis 2-63: Preparation of(2R,3R,4R,5R,6S)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A91)

Step 1: To a stirred solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-nitro-3,4-dihydro-2H-pyran(A91-1, 1.0 g, 2.17 mmol) in dry THF (10 ml) was added NaOMe (0.65 mL, 5M in MeOH). After stirring at rt for 1 h, the reaction mixture wasneutralized with Amberlite IR-120 resin (H⁺). The reaction mixture wasfiltered, the filtrate was concentrated to give a crude product, whichwas purified by column to get((2R,3R,4R,5R,6S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-6-methoxy-5-nitrotetrahydro-2H-pyran(A91-2, 425 mg, 39% yield) and(2R,3R,4R,5R,6R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-6-methoxy-5-nitrotetrahydro-2H-pyran(52 mg, 5%). LC-MS (ESI) of both found: 494 [M+H]⁺.

Step 2: To a solution of(2R,3R,4R,5R,6S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-6-methoxy-5-nitrotetrahydro-2H-pyran(A91-2, 425 mg, 0.86 mmol) in MeOH (10 mL) was added Raney Ni (50 mg).The mixture was stirred at rt for 12 h under a H₂ balloon. The mixturewas filtered and concentrated to give(2S,3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A91-3, 473 mg, 72% yield). LC-MS (ESI) found: 464 [M+H]⁺.

Step 3: To a solution of(2S,3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A91-3, 200 mg, 0.431 mmol) in MeOH (10 mL) was added Pd/C (20 mg, 10%wt, 60% wet) and several drops of 1 N HCl. The mixture was stirred at rtfor 12 h under a H₂ balloon. The mixture was filtered and concentratedto give(2R,3R,4R,5R,6S)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A91, 452 mg, 62% yield) as colorless oil. LC-MS (ESI) found: 194[M+H]⁺.

Alternatively,(2R,3R,4R,5R,6S)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A91) can be synthesized in the following manner:

To a solution ofN-[(2S,3R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxyoxan-3-yl]acetamide(A91-4, 2 g, 8.5 mmol) in H₂O (11 mL) was added Ba(OH)₂ (9.5 g, 55.3mmol). The mixture was heated for reflux at 120° C. overnight. Later thesolution of (NH₄)₂SO₄ (7.0 g, 55.3 mmol) in H₂O (55 mL) was slowly addedinto the mixture. The mixture was heated to reflux at 120° C. for fullyreaction. After cooling down to room temperature, the mixture wasfiltered and the filtrate was concentrated. The residual was thenadjusted to pH=7 by adding MeONa through the environment of MeOH. Thesolution was concentrated and recrystallized in i-PrOH to give(2R,3R,4R,5R,6S)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A91, 1.6 g, 97% yield) as a white solid. LC-MS (ESI) found: 194 [M+H]⁺.¹H NMR (400 MHz, CD₃OD): δ4.71 (d, J=3.7 Hz, 1H), 3.83 (d, J=3.2 Hz,1H), 3.78-3.66 (m, 3H), 3.54 (dd, J=10.4, 3.2 Hz, 1H), 3.40 (s, 3H),2.96 (dd, J=10.4, 3.7 Hz, 1H).

Synthesis 2-64: Preparation of(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A92)

Step 1: To a mixture of(2S,3R,4R,5R,6R)-3-acetamido-6-(acetoxymethyl)tetrahydro-2H-pyran-2,4,5-triyltriacetate (A92-1, 20.0 g, 51.4 mmol) in DCM (200 mL) was added TiCl₄(61.6 mL, 1 M in DCM) at 0° C. under N₂. After refluxing at 50° C.overnight, the mixture was concentrated and purified by chromatography(0-80% ethyl acetate in petroleum) to give(2R,3R,4R,5R)-5-acetamido-2-(acetoxymethyl)-6-chlorotetrahydro-2H-pyran-3,4-diyldiacetate (A92-2, 8.5 g, 45% yield) as white solid. LC-MS (ESI) found:366 [M+1]⁺.

Step 2: To a mixture of(2R,3R,4R,5R)-5-acetamido-2-(acetoxymethyl)-6-chlorotetrahydro-2H-pyran-3,4-diyldiacetate (A92-2, 8.5 g, 23.2 mmol) in toluene (85 mL) was added Bu₃SnH(8.1 g, 27.9 mmol) and AIBN (0.08 g, 0.46 mmol) at rt under N₂. Afterrefluxing at 110° C. for 1.5 h, the mixture was concentrated andpurified by chromatography on silica gel (70-100% ethyl acetate inpetroleum) to give(2R,3R,4R,5S)-5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-3,4-diyldiacetate (A92-3, 6.6 g, 86%) as white solid. LC-MS (ESI) found: 332[M+1]⁺.

Step 3: To a mixture of(2R,3R,4R,5S)-5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-3,4-diyldiacetate (A92-3, 6.6 g, 19.9 mmol) in H₂O (48 mL) was added HCl (12 mL,2.5 M in H₂O) at rt under N₂. After refluxing at 100° C. for 2 h, themixture was concentrated. EtOH (10 mL) and Et₂O (10 mL) were added. Thesolid formed was filtered to give(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diolhydrochloride (A-92, 2.7 g, 68%) as white solid. LC-MS (ESI) found: 164[M+H]⁺.

Synthesis 2-65: Preparation of(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(Compound A94)

Step 1: To a mixture of(2R,3R,4R,5S)-5-acetamido-2-(acetoxymethyl)tetrahydro-2H-pyran-3,4-diyldiacetate (A94-1, 1.5 g, 4.5 mmol) in MeOH (20 mL) was added NaOMe (2.7mL, 5 M in MeOH) at 0° C. under N₂. After stirring at rt for 2 h, themixture was neutralized with HCl (2 M) and concentrated. Then themixture was purified by chromatography on silica gel (0-20% methanol inDCM) to giveN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(A94-2, 560 mg, 60%) as white solid. LC-MS (ESI) found: 206 [M+H]⁺.

Step 2: To a mixture ofN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(A-94-2, 130 mg, 0.63 mmol) in DMF (5.0 mL) was added NaH (101 mg, 4.2mmol) at 0° C. under N₂. After stirring at 0° C. for 30 min, BnBr (0.07mL, 0.63 mmol) was added slowly and the reaction was stirred for another1 hour. The mixture was quenched with H₂O and extracted with EA. Thecombined organic layers were washed with brine, dried over anhydrousNa₂SO₄. The residue was concentrated and purified by chromatography onsilica gel (0-20% methanol in DCM) to giveN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(A94-3, 130 mg, 43%) as white solid. LC-MS (ESI) found: 476 [M+H]⁺.

Step 3: To a mixture ofN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(A94-3, 1.2 g, 2.5 mmol) in THE (15 mL) was added Et₃N (1.1 mL, 7.6mmol), DMAP (30 mg, 0.25 mmol) and Boc₂O (7.0 mL, 30.2 mmol) at 0° C.under N₂. After stirring at rt overnight, the mixture was concentratedand purified by chromatography on silica gel (0-10% methanol in DCM) togive tert-butylacetyl((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A94-4, 860 mg, 60%) as colorless oil. LC-MS (ESI) found: 576 [M+1]⁺.

Step 4: To a mixture of tert-butylacetyl((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A94-4, 860 mg, 1.5 mmol) in THE (10 mL) was added 2 mL NaOH (40% aq)under N₂. After refluxing at 60° C. overnight, the mixture was dilutedwith H₂O and extracted with EA. The combined organic layer was washedwith brine, dried over Na₂SO₄, filtered. The filtrate was concentratedand purified by chromatography on silica gel (0-70% ethyl acetate inpetroleum) to give tert-butyl((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A94-5, 450 mg, 56%) as a white solid. LC-MS (ESI) found: 534 [M+1]⁺.

Step 5: To a solution of tert-butyl((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A94-5, 450 mg, 0.84 mmol) in DCM (9.0 mL) was added TFA (3.0 mL) at rtunder N₂. After stirring for 2 h, the mixture was quenched withNaHCO₃(aq) and extracted with EA. The combined organic layer was washedwith brine, dried over anhydrous Na₂SO₄. The residue was concentratedand purified by chromatography on silica gel (0-50% ethyl acetate inpetroleum) to give(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A94, 280 mg, 77%) as colorless oil. LC-MS (ESI) found: 434 [M+1]⁺.

Synthesis 2-66: Preparation of4-chloro-2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)thiazole-5-carbonitrile(Compound A95)

To a mixture of(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diolhydrochloride (A92, 50 mg, 0.25 mmol) in NMP (2.0 mL) was added2,4-dichlorothiazole-5-carbonitrile (135 mg, 0.75 mmol) and DIPEA (0.17mL, 1.0 mmol) at rt under N₂. After stirring at 120° C. overnight, themixture was concentrated and purified by prep-TLC to give4-chloro-2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)thiazole-5-carbonitrile(A95, 6.3 mg, 8% yield) as brown solid. LC-MS (ESI) found: 306 [M+H]⁺.¹H NMR (400 MHz, CD3OD): δ4.12-4.00 (m, 2H), 3.90 (d, J=2.5 Hz, 1H),3.77-3.65 (m, 2H), 3.58 (dd, J=10.1, 3.2 Hz, 1H), 3.46-3.41 (m, 1H),3.16 (t, J=10.5 Hz, 1H).Synthesis 2-67: Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-(pyridin-2-ylamino)tetrahydro-2H-pyran-3,4-diol(Compound A96)

Step 1: To a solution of(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A-94, 40.0 mg, 0.092 mmol) in toluene (4 mL) was added 2-bromopyridine(21.9 mg, 0.139 mmol), Pd(OAc)₂ (2.07 mg, 0.009 mmol), BINAP (5.75 mg,0.009 mmol), and NaOt-Bu (26.6 mg, 0.277 mmol). The mixture was stirredunder N₂ at 100° C. for 24 h. The reaction mixture was concentrated andpurified by prep-HPLC (Method A) to giveN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)pyridin-2-amine(A96-1, 15 mg, 32%) as white solid. LC-MS (ESI) found: 511 [M+H]⁺.

Step 2: To a solution ofN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)pyridin-2-amine(A96-1, 15 mg, 0.03 mmol) in MeOH (5 mL) was added Pd/C (2 mg, 10% wt,60% wet). The mixture was stirred at rt for 12 h under a H₂ balloon. Themixture was filtered and the filtrate was concentrated to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-(pyridin-2-ylamino)tetrahydro-2H-pyran-3,4-diol(A96, 0.6 mg, 9% yield) as white solid. LC-MS (ESI) found: 241 [M+H]⁺.¹H NMR (400 MHz, Methanol-d₄): δ 8.03-7.81 (m, 1H), 7.42 (ddd, J=8.8,7.0, 2.0 Hz, 1H), 6.67-6.38 (m, 2H), 4.67-4.37 (m, 1H), 4.16-3.96 (m,1H), 3.89 (d, J=3.1 Hz, 1H), 3.79-3.58 (m, 2H), 3.54 (dd, J=9.9, 3.0 Hz,1H), 3.44 (t, J=6.0 Hz, 1H), 3.21-2.99 (m, 1H).

The following compounds below were made using the method described inSynthesis 2-66 or Synthesis 2-67:

A97

A98

A99

A100

A101

A102

A103

A104

ID Characterization data A97  LC-MS (ESI) found: 260 [M+H]⁺. ¹H NMR (400MHz, Methanol-d₄): δ 8.25 (d, J = 2.2 Hz, 1H), 8.10-7.91 (m, 1H), 4.59(td, J = 10.7, 5.3 Hz, 1H), 4.05 (dd, J = 11.0, 5.3 Hz, 1H), 3.92 (dd, J= 3.3, 1.0 Hz, 1H), 3.81- 3.65 (m, 3H), 3.46 (ddd, J = 6.6, 5.0, 1.1 Hz,1H), 3.21 (t, J = 10.9 Hz, 1H). A98  Yield: 3.0 mg, 12%, white solid.LC-MS (ESI) found: 242 [M + H]⁺. ¹H NMR (400 MHz, Methanol-d₄): δ 8.37(s, 1H), 7.99 (d, J = 5.9 Hz, 1H), 6.54 (d, J = 5.4 Hz, 1H), 4.66-4.49(m, 1H), 4.07 (dd, J = 9.8, 5.0 Hz, 1H), 3.90 (d, J = 1.8 Hz, 1H),3.80-3.73 (m, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.59 (dd, J = 10.4,3.1 Hz, 1H), 3.44 (ddd, J = 6.9, 5.1, 1.2 Hz, 1H), 3.12 (t, J = 10.9 Hz,1H). A99  Yield: 2.2 mg, 3.1%, yellow solid. LC-MS (ESI) found: 281 [M +H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 6.33 (s, 1H), 4.11 (dd, J = 11.1, 5.2Hz, 1H), 3.93 (dt, J = 10.5, 5.2 Hz, 1H), 3.88 (d, J = 3.0 Hz, 1H),3.77-3.65 (m, 2H), 3.54 (dd, J = 10.3, 3.2 Hz, 1H), 3.44-3.41 (m, 1H),3.13 (t, J = 10.9 Hz, 1H). A100 Yield: 32 mg, 20%, white solid. LC-MS(ESI) found: 293 [M + H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 7.79 (d, J = 2.1Hz, 1H), 7.34 (dd, J = 10.7, 2.1 Hz, 1H), 4.41 (td, J = 10.6, 5.2 Hz,1H), 4.10 (dd, J = 11.0, 5.2 Hz, 1H), 3.90 (d, J = 2.8 Hz, 1H), 3.76(dd, J = 11.3, 7.0 Hz, 1H), 3.71-3.65 (m, 2H), 3.47-3.43 (m, 1H), 3.15(t, J = 10.8 Hz, 1H). A101 Yield: 2.8 mg, 9%, white solid. LC-MS (ESI)found: 259 [M + H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 7.46 (dd, J = 16.3, 8.1Hz, 1H), 6.37 (dd, J = 8.1, 2.2 Hz, 1H), 6.07 (dd, J = 7.7, 2.0 Hz, 1H),4.21-4.08 (m, 2H), 3.90 (d, J = 2.7 Hz, 1H), 3.76 (dd, J = 11.4, 7.1 Hz,1H), 3.69 (dd, J = 11.4, 5.0 Hz, 1H), 3.56 (dd, J = 10.3, 3.2 Hz, 1H),3.46-3.41 (m, J = 6.9, 5.0, 0.9 Hz, 1H), 3.08 (t, J = 10.7 Hz, 1H). ¹⁹FNMR (377 MHz, CD3OD): δ- 73.05 (s). A102 Yield: 11 mg, 17%, yellowsolid. LC-MS (ESI) found: 248 [M + H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 7.85(s, 1H), 4.13 (dd, J = 11.1, 5.2 Hz, 1H), 3.92 (dd, J = 21.5, 3.7 Hz,2H), 3.78-3.66 (m, 2H), 3.61 (dd, J = 10.3, 3.2 Hz, 1H), 3.45 (ddd, J =6.9, 5.0, 1.0 Hz, 1H), 3.19 (t, J = 10.9 Hz, 1H). A103 Yield: 3.6 mg,4%, white solid. LC-MS (ESI) found: 277 [M + H]⁺. ¹H NMR (400 MHz,CD₃OD): δ 7.16-7.05 (m, 2H), 3.98 (dd, J = 11.3, 5.1 Hz, 1H), 3.91 (d, J= 2.5 Hz, 1H), 3.80-3.72 (m, 2H), 3.69 (dd, J = 11.4, 5.0 Hz, 1H), 3.61(dd, J = 10.0, 3.2 Hz, 1H), 3.45 (ddd, J = 6.9, 5.0, 0.9 Hz, 1H), 3.18(t, J = 11.0 Hz, 1H). A104 Yield: 7 mg, 10%, white solid. LC-MS (ESI)found: 282 [M + H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 4.12 (dd, J = 11.1, 5.1Hz, 1H), 3.90 (d, J = 2.6 Hz, 2H), 3.71 (dt, J = 11.4, 5.4 Hz, 2H), 3.60(dd, J = 10.3, 3.2 Hz, 1H), 3.46-3.43 (m, 1H), 3.19 (t, J = 10.9 Hz,1H).Synthesis 2-68: Preparation of(2R,3R,4R,5R,6S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-211-pyran-3,4-diol(Compound A105) and(2R,3R,4R,5R,6R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H1-pyran-3,4-diol(Compound Al06)

A solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 224.0 mg, 1.2 mmol), 3,5-dichloro-1,2,4-thiadiazole (372.0 mg, 2.4mmol) and DIEA (464.4 mg, 3.6 mmol) in i-PrOH (10 mL) was stirred at rtovernight. The mixture was concentrated in vacuo. The crude product waspurified by flash chromatography (silica gel, 0-10% MeOH in DCM) andpre-HPLC (Method B) to give(2R,3R,4R,5R,6S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(0.8 mg, 0.2% yield) as a white solid and(2R,3R,4R,5R,6R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(3.7 mg, 1% yield) as white solid.

Compound A105: LC-MS (ESI) found: 312 [M+H]⁺. ¹H NMR (400 MHz,Methanol-d₄): δ 4.85-4.80 (m, 1H), 4.23-4.11 (m, 1H), 3.91 (d, J=3.1 Hz,1H), 3.85-3.77 (m, 2H), 3.76-3.69 (m, 2H), 3.40 (s, 3H). Compound A106:LC-MS (ESI) found: 312 [M+H]⁺. ¹H NMR (400 MHz, Methanol-d₄): δ 4.33 (d,J=8.1 Hz, 1H), 3.87 (d, J=3.2 Hz, 1H), 3.84-3.70 (m, 2H), 3.68 (dd,J=10.4, 3.3 Hz, 1H), 3.53 (ddd, J=6.7, 5.4, 1.1 Hz, 1H), 3.48 (s, 3H),3.46-3.40 (m, 1H).

Synthesis 2-69: Preparation of(2R,3R,4R,5R,6R)-5-((6-fluoropyridin-2-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A107)

To a solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 50.0 mg, 0.259 mmol) in NMP (2 mL) was added 2,6-difluoropyridine(89.0 mg, 20 0.777 mmol) and DIPEA (101 mg, 0.777 mmol). The mixture wasstirred at 180° C. for 1 h under microwave. The reaction mixture waslyophilized and purified by prep-HPLC (Method A) to give(2R,3R,4R,5R,6R)-5-((6-fluoropyridin-2-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A107, 1.4 mg, 2% yield) as white solid. LC-MS (ESI) found: 289 [M+H]⁺.¹H NMR (400 MHz, Methanol-d₄): δ 7.47 (q, J=8.2 Hz, 1H), 6.43 (dd,J=8.2, 2.3 Hz, 1H), 6.07 (dd, J=7.7, 2.0 Hz, 1H), 4.31 (d, J=8.2 Hz,1H), 3.95-3.84 (m, 2H), 3.77 (h, J=4.9 Hz, 2H), 3.62 (dd, J=10.4, 3.3Hz, 1H), 3.53 (ddd, J=6.7, 5.4, 1.1 Hz, 1H), 3.45 (s, 3H).

The following compounds below were made using the method described inSynthesis 2-68 or Synthesis 2-69:

A108

A109

A110

A111

ID Characterization data Starting Material A111 LC-MS (ESI) found: 336[M + H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 4.30 (d, J = 7.8 Hz, 1H), 3.86 (d,J = 2.1 Hz, 1H), 3.81-3.71 (m, 2H), 3.64 (dt, J = 14.5, 7.3 Hz, 2H),3.54-3.50 (m, 1H), 3.48 (s, 3H).

A110 LC-MS (ESI) found: 277 [M + H]⁺. ¹H NMR (400 MHz, CD₃OD): δ 7.82(s, 1H), 4.34 (d, J = 8.1 Hz, 1H), 3.87 (d, J = 3.1 Hz, 1H), 3.83-3.68(m, 4H), 3.55-3.51 (m, 1H), 3.47 (s, 3H).

A109 Yield: 2 mg, 6%, white solid. LC-MS (ESI) found: 323 [M + H]⁺. ¹HNMR (400 MHz, CD₃OD): δ 7.76 (d, J = 2.1 Hz, 1H), 7.34 (dd, J = 10.8,2.0 Hz, 1H), 4.44 (d, J = 8.4 Hz, 1H), 4.32-4.26 (m, 1H), 3.88 (d, J =3.1 Hz, 1H), 3.84-3.76 (m, 2H), 3.73 (dd, J = 10.6, 3.3 Hz, 1H), 3.54(t, J = 6.2 Hz, 1H), 3.45 (s, 3H). ¹⁹F NMR (377 MHz, CD₃OD): δ-139.12(s).

A108 Yield: 0.5 mg, 2%, white solid. LC-MS (ESI) found: 311 [M + H]⁺. ¹HNMR (400 MHz, Methanol-d₄): δ 6.31 (s, 1H), 4.29 (d, J = 8.0 Hz, 1H),3.86 (d, J = 3.2 Hz, 1H), 3.77 (t, J = 6.1 Hz, 2H), 3.63 (dd, J = 10.3,3.2 Hz, 1H), 3.56 (d, J = 8.0 Hz, 1H), 3.52 (d, J = 6.1 Hz, 1H), 3.47(s, 3H).

Preparation of(2R,3R,4R,5R,6R)-5-((1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A110)

Step 1: A solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(30 mg, 0.16 mmol), DIPEA (40 mg, 0.31 mmol) and5-chloro-1,2,4-thiadiazole (22.4 mg, 0.19 mmol) in i-PrOH (1 mL) wasstirred at 120° C. overnight. The mixture was concentrated and theresidue was purified by prep-HPLC (Method A) to give(2R,3R,4R,5R,6R)-5-((1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(6.8 mg, 16% yield) as a white solid.

Synthesis 2-70: Preparation of6-(((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)amino)-1,3,5-triazine-2,4(1H,3H)-dione(Compound A112)

To a solution of 2,4,6-trichloro-1,3,5-triazine (187 mg 1.03 mmol) inTHF (5 mL) was added DIPEA (200 mg, 1.55 mmol) at −78° C.(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 100 mg, 0.51 mmol) was then added at −78° C. The mixture wasfurther stirred for 2 h at −78° C. Then it was quenched by adding H₂O (5mL). The mixture was warmed to rt and stirred for another 2 h. Thesolvent was evaporated and the residual was purified by prep-HPLC(Method A) to give6-(((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)amino)-1,3,5-triazine-2,4(1H,3H)-dione(A112, 6.1 mg, 4% yield) as white solid. LC-MS (ESI) found: 305 [M+H]⁺.¹H NMR (400 MHz, D₂O): δ4.39 (d, J=8.4 Hz, 1H), 4.02 (dd, J=10.7, 8.5Hz, 1H), 3.88 (d, J=3.2 Hz, 1H), 3.81-3.67 (m, 3H), 3.63 (dd, J=7.8, 4.3Hz, 1H).

The following compound below was made using the method described inSynthesis 2-70 with(2R,3R,4R,5R,6S)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diolinstead of A90:

A113

ID Characterization data A113 LC-MS (ESI) found: 323 [M + H]⁺. ¹H NMR(400 MHz, CD₃OD): δ 8.51 (s, 1H), 7.79 (d, J = 2.1 Hz, 1H), 7.37 (dd, J= 10.7, 2.1 Hz, 1H), 4.83 (s, 1H), 4.56 (d, J = 3.7 Hz, 1H), 3.93 (d, J= 3.1 Hz, 1H), 3.86 (dd, J = 10.8, 3.2 Hz, 1H), 3.82-3.79 (m, 1H),3.77-3.71 (m, 2H), 3.37 (s, 3H).Synthesis 2-71: Preparation of(2R,3R,4R,5R,6S)-5-((3-(dimethylamino)-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A114)

A solution of(2R,3R,4R,5R,6S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A105, 10.0 mg, 0.03 mmol), dimethylamine (0.1 mL, 0.1 mmol, 1 M in THF)and DIEA (11.6 mg, 0.09 mmol) in NMP (4 mL) was stirred at 120° C.overnight. The mixture was concentrated in vacuo. The crude product waspurified by pre-HPLC (Method B) to give(2R,3R,4R,5R,6S)-5-((3-(dimethylamino)-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A114, 0.8 mg, 8.3% yield) as white solid. LC-MS (ESI) found: 321[M+H]⁺. ¹H NMR (400 MHz, Methanol-d₄): δ 4.85-4.81 (m, 1H), 4.12 (d,J=13.7 Hz, 1H), 3.90 (d, J=3.2 Hz, 1H), 3.81 (ddd, J=11.4, 7.8, 3.3 Hz,2H), 3.77-3.67 (m, 2H), 3.39 (s, 3H), 3.04 (s, 6H).The following compounds below were made using the method described inSynthesis 2-71 with the appropriate amine and, A106 instead of A105:

Preparation of(2R,3R,4R,5R,6R)-2-(hydroxymethyl)-6-methoxy-5-((3-(4-methylpiperazin-1-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A116)

Step 1: A solution of(2R,3R,4R,5R,6R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(10 mg, 0.03 mmol), 1-methylpiperazine (5.0 mg, 0.05 mmol) and DIEA(11.6 mg, 0.09 mmol) in i-PrOH (4 mL) was stirred at 120° C. overnight.The mixture was concentrated in vacuo. The crude product was purified byprep-HPLC (Method A) to give product (1.4 mg, 7% yield) as white solid.LC-MS (ESI) found: 376 [M+H]⁺. ¹H NMR (400 MHz, Methanol-d₄): δ 4.35 (d,J=8.2 Hz, 1H), 3.86 (d, J=3.2 Hz, 1H), 3.81-3.74 (m, 2H), 3.70 (dd,J=10.5, 3.4 Hz, 1H), 3.57 (t, J=5.1 Hz, 4H), 3.54-3.50 (m, 2H), 3.48 (s,3H), 2.48 (t, J=5.1 Hz, 4H), 2.31 (s, 3H).

Preparation of(2R,3R,4R,5R,6R)-2-(hydroxymethyl)-6-methoxy-5-((3-morpholino-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A117)

It was prepared according to the procedure same as that for A116. Yield:1.0 mg, 9%, white solid. LC-MS (ESI) found: 363 [M+H]⁺. ¹H NMR (400 MHz,Methanol-d₄): δ 4.35 (d, J=8.2 Hz, 1H), 3.86 (d, J=3.2 Hz, 1H), 3.77 (t,J=6.2 Hz, 2H), 3.73-3.69 (m, 6H), 3.54-3.45 (m, 8H).

ID Characterization data A115 LC-MS (ESI) found: 321 [M + H]⁺. ¹H NMR(400 MHz, CD₃OD): δ 4.36 (d, J = 8.2 Hz, 1H), 3.86 (d, J = 3.1 Hz, 1H),3.81-3.69 (m, 3H), 3.55-3.50 (m, 1H), 3.48 (s, 3H), 3.46-3.40 (m, 1H),3.03 (s, 6H). A116 LC-MS (ESI) found: 376 [M + H]⁺. ¹H NMR (400 MHz,CD3OD) δ: 4.35 (d, J = 8.2 Hz, 1H), 3.86 (d, J = 3.2 Hz, 1H), 3.81-3.74(m, 2H), 3.70 (dd, J = 10.5, 3.4 Hz, 1H), 3.57 (t, J = 5.1 Hz, 4H),3.54-3.50 (m, 2H), 3.48 (s, 3H), 2.48 (t, J = 5.1 Hz, 4H), 2.31 (s, 3H).A117 LC-MS (ESI) found: 363 [M + H]⁺. ¹H NMR (400 MHz, Methanol-d₄): δ4.35 (d, J = 8.2 Hz, 1H), 3.86 (d, J = 3.2 Hz, 1H), 3.77 (t, J = 6.2 Hz,2H), 3.73-3.69 (m, 6H), 3.54-3.45 (m, 8H).Synthesis 2-72: Preparation ofN-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)acrylamide(Compound A118)

Step 1: A solution of compound(2R,3R,4R,5R,6R)-5-azido-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A89, 1 g, 4.6 mmol) in anhydrous pyridine (20 mL) was treated withtrimethylsilyl chloride (3.5 mL, 27.8 mmol) and the mixture was stirredfor 12 h at room temperature. The solvent was evaporated and the residuewas diluted in ethyl acetate/water. The organic layer was separated andfurther washed by water, brine, dried over anhydrous MgSO₄, filtered,and concentrated to afford desired product as a yellow oil (1.6 g, 81%yield). LC-MS (ESI) found: 436 [M+H]⁺.

Step 2: To a solution of(((2R,3S,4R,5R,6R)-5-azido-6-methoxy-2-(((trimethylsilyl)oxy)methyl)tetrahydro-2H-pyran-3,4-diyl)bis(oxy))bis(trimethylsilane)(A118-1, 1.0 g, 2.29 mmol) in MeOH (20 mL) was added Pd/C (100 mg, 10%wt, 60% wet). The mixture was stirred at rt for 12 h under a H₂ balloon.The mixture was filtered and concentrated to give((2R,3S,4R,5R,6R)-5-amino-6-methoxy-3,4-bis((trimethylsilyl)oxy)tetrahydro-2H-pyran-2-yl)methanol(A118-2, 401 mg, 52% yield). LC-MS (ESI) found: 338 [M+H]⁺.

Step 3: To a solution of((2R,3S,4R,5R,6R)-5-amino-6-methoxy-3,4-bis((trimethylsilyl)oxy)tetrahydro-2H-pyran-2-yl)methanol(A118-2, 150 mg, 0.444 mmol) in MeOH (1 mL) H₂O (1 mL) was addedprop-2-enoyl chloride (0.04 mL, 0.533 mmol) and TEA (0.02 mL, 0.148mmol). The mixture was stirred at 0° C. for 4 h. The solvent wasevaporated and the residue was diluted in DCM/water. The organic layerwas separated and further washed by water, brine, dried over anhydrousMgSO₄, filtered, and concentrated to afford a crude product, which waspurified by silica gel column to giveN-((2R,3R,4R,5S,6R)-6-(hydroxymethyl)-2-methoxy-4,5-bis((trimethylsilyl)oxy)tetrahydro-2H-pyran-3-yl)acrylamide(A118-3, 70 mg, 40%). LC-MS (ESI) found: 392 [M+H]⁺.

Step 4: To a solution ofN-((2R,3R,4R,5S,6R)-6-(hydroxymethyl)-2-methoxy-4,5-bis((trimethylsilyl)oxy)tetrahydro-2H-pyran-3-yl)acrylamide(A118-3, 70 mg, 0.179 mmol) in 5 THE (1 mL) was added TBAF (0.2 mL, 1 Min THF). The mixture was stirred at 0° C. for 30 min.

The solvent was evaporated and the residual was purified by prep-HPLC(Method A) to giveN-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)acrylamide(A118, 17 mg, 38%) as white solid. LC-MS (ESI) found: 248 [M+H]⁺. ¹H NMR(400 MHz, Methanol-d₄): δ 6.04-6.17 (m, 2H), 5.63-5.66 (m, 1H), 4.29 (d,J=8.4 Hz, 1H), 3.79-3.83 (m, 2H), 3.55-3.65 (m, 4H), 3.35 (s, 3H).

Synthesis 2-73: Preparation of1-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)pyridin-4(1H)-one(Compound A119)

To a solution of 4H-pyran-4-one (9.6 mg, 0.100 mmol) in MeOH (3 mL) wasadded NaOH (8 mg, 0.2 mmol) in H₂O (2 mL) to adjust pH to 11, then(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 20 mg, 0.1 mmol) was added to the mixture. The mixture was stirredat 60° C. for 3 h. Then the solvent was evaporated and the residual waspurified by prep-HPLC (Method A) to give1-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)pyridin-4(1H)-one(A119, 2.1 mg, 7.7%) as white solid. LC-MS (ESI) found: 272 [M+H]⁺. ¹HNMR (400 MHz, Methanol-d₄): δ 8.49 (s, 1H), 7.86 (d, J=7.6 Hz, 2H), 6.46(d, J=7.6 Hz, 2H), 4.65 (d, J=8.3 Hz, 1H), 4.08 (dd, J=10.9, 3.3 Hz,1H), 3.98-3.89 (m, 2H), 3.85-3.75 (m, 2H), 3.68 (t, J=6.1 Hz, 1H), 3.42(s, 3H).Synthesis 2-74: Preparation ofN-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)-4-methylbenzenesulfonamide(Compound A120)

A solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 50 mg, 0.26 mmol) in MeOH (0.5 mL) and H₂O(0.5 mL) was added DIPEA(0.92 mL, 5.2 mmol) and 4-methylbenzene-1-sulfonyl chloride (0.25 mL,1.3 mmol). After stirring under N₂ at rt overnight, the mixture wasconcentrated to give crude product which was further purified byprep-HPLC (Method B) to affordN-((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)-4-methylbenzenesulfonamide(A120, 7.0 mg, 15%) as white solid. LC-MS (ESI) found: 348 [M+1]⁺. ¹HNMR (400 MHz, DMSO-d₆): δ 7.64 (d, J=8.3 Hz, 2H), 7.48 (s, 1H), 7.31 (d,J=8.0 Hz, 2H), 4.55 (dd, J=7.7, 3.6 Hz, 2H), 4.50 (d, J=6.6 Hz, 1H),3.88 (d, J=8.1 Hz, 1H), 3.63 (t, J=3.6 Hz, 1H), 3.53-3.40 (m, 2H), 3.30(dd, J=6.7, 3.2 Hz, 1H), 3.22 (dd, J=8.0, 4.2 Hz, 2H), 2.85 (s, 3H),2.36 (s, 3H).Synthesis 2-75: Preparation ofN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-4-methylbenzenesulfonamide(Compound A121)

Step 1: To a mixture of(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A94, 50 mg, 0.12 mmol) in MeOH (1.0 mL) and H₂O (1.0 mL) was added TsCl(219 mg, 1.2 mmol) and TEA (175 mg, 1.7 mmol) at 0° C. under N₂. Afterstring for 2 h, the mixture was concentrated and purified by prep-HPLC(Method A) to giveN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-4-methylbenzenesulfonamide(A121-1, 30 mg, 44%) as a white solid. LC-MS (ESI) found: 588 [M+1]⁺.

Step 2: To a mixture ofN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-4-methylbenzenesulfonamide(A121-1, 30 mg, 0.051 mmol) in MeOH (3.0 mL) was added Pd/C (10 mg, 10%wt, 60% wet) at rt. The mixture was stirred at rt for 12 h under a H₂balloon. The mixture was filtered and concentrated to affordN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-4-methylbenzenesulfonamide(A121, 10 mg, 62%) as white solid. LC-MS (ESI) found: 318 [M+1]⁺. ¹H NMR(400 MHz, CD3OD): δ7.78 (d, J=8.3 Hz, 2H), 7.36 (d, J=8.0 Hz, 2H), 3.80(dd, J=2.9, 0.8 Hz, 1H), 3.75 (dd, J=11.2, 4.8 Hz, 1H), 3.62 (ddd,J=16.4, 11.4, 6.0 Hz, 2H), 3.45-3.31 (m, 3H), 3.08-3.00 (m, 1H), 2.42(s, 3H).

Synthesis 2-7: Preparation of(3R,4R,5R,6R)-3-(4-fluorophenyl)-6-(hydroxymethyl)tetrahydro-2H-pyran-2,4,5-triol(Compound A122)

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(A122-1, 100 mg, 0.184 mmol) in DME (10 mL) was added Pd(PPh₃)₄ (21 mg,0.018 mmol), K₂CO₃ (76 mg, 0.552 mmol) and (4-fluorophenyl)boronic acid(34 mg, 0.239 mmol). The mixture was charged with N₂ for three times andstirred at 90° C. under N₂ for 16 h. The mixture was filtered, thefiltrate was concentrated under reduced pressure to give a crudeproduct, which was purified by flash chromatography (silica gel, 5-10%EtOAc in PE) to give(2R,3R,4R)-4-(benzyloxy)-2-((benzyloxy)methyl)-5-(4-fluorophenyl)-3,4-dihydro-2H-pyran-3-ol(A122-2, 25 mg, 31% yield) as colorless oil. LC-MS (ESI) found: 533[M+23]⁺.

(2R,3R,4R,5R)-5-(2,4-difluorophenyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A122a),(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1H-pyrazol-3-yl)tetrahydro-2H-pyran-3,4-diol(Compound A122b) and(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1H-pyrazol-4-yl)tetrahydro-2H-pyran-3,4-diol(Compound A122c) were prepared using the procedure shown in Synthesis2-7

Alternatively,(2R,3R,4R,5R)-5-(2,4-difluorophenyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A122a) can be synthesized in the following manner:

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(433 mg, 0.80 mmol) in DME (30 mL) and water (10 mL) were added(2,4-difluorophenyl)boronic acid (164 mg, 1.0 mmol) and K₂CO₃(331 mg,2.4 mmol). The mixture was stirred at 90° C. under N₂ for 16 h. Themixture was concentrated under reduced pressure to give a crude product,which was purified by silica gel column to give(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2,4-difluorophenyl)-3,4-dihydro-2H-pyran(200 mg, 47% yield). LC-MS (ESI) found: 529 [M+H]⁺.

Step 2: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(2,4-difluorophenyl)-3,4-dihydro-2H-pyran(30 mg, 0.057 mmol) in MeOH (10 mL) was added Pd/C (5 mg, 10% wt, 60%wet), the mixture was stirred at rt overnight under a H₂ balloon. Themixture was filtered, the filtrate was concentrated to give(2R,3R,4R,5R)-5-(2,4-difluorophenyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(9.7 mg, 65% yield). LC-MS (ESI) found: 261 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 7.79 (m, 1H), 6.84 (m, 2H), 4.20 (dd, J=11.7, 6.9 Hz, 1H), 4.09(dd, J=13.0, 8.9 Hz, 1H), 4.04-3.95 (m, 1H), 3.92-3.86 (m, 1H), 3.76 (m,2H), 3.66 (dd, J=11.7, 4.1 Hz, 1H), 3.33 (dd, J=7.3, 3.9 Hz, 1H).

Example 3. Synthesis of Degraders Synthesis 3-1. Preparation ofBidentate Fumaramide OPT-3 (Compound 1)

The —NH(OPT-3)C(O)CH₂SH is generated in situ from NH₂OH treatment ofSATA-(N-succinimidyl S-acetylthioacetate)-OPT-3.

Synthesis 3-2. General Synthesis of Bidentate Fumaramide OPT-3conjugate-sulfoximine Compounds

The —NH(OPT-3)C(O)CH₂SH is generated in situ from NH₂OH treatment ofSATA-(N-succinimidyl S-acetylthioacetate)-OPT-3. Compound 3 can besynthesized using the procedure of Scheme 2-59 with MeLi.

Synthesis 3-3. Preparation of Compound 4

Compound 4-1 is synthesized from ASGPR Ligand A41 from Synthesis 2-33

Synthesis 3-4. Preparation of Compound 5

Compound 5-1 is synthesized from ASGPR Ligand A51 from Synthesis 2-39

Synthesis 3-5. Preparation of Compound 6 and Compound 7

Synthesis 3-6. Preparation of Compound 8

Synthesis 3-7. Preparation of Compound 9

Example 4. Synthesis of Talose-Based ASGPR ligands

Synthesis 4-1. Preparation of Bicyclic Talose amine Compound A123,Compound A124, and Compound A125:

Synthesis 4-2. Preparation of tert-Butyl(((3aR,4S,8S,8aR)-8-amino-2,2-dimethyltetrahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4(5H)-yl)methyl)carbamate(Intermediate 1b):

Synthesis 4-3. Preparation of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran(Compound A126)

Step 1: To a solution of(2R,3R,4R)-2-(acetoxymethyl)-3,4-dihydro-2H-pyran-3,4-diyl diacetate(A126-1, 30.0 g, 55.1 mmol) in MeOH (300 mL) was added NaOMe (31.0 mL,165.3 mmol, 5.4 M in MeOH) at room temperature. The mixture was stirredat room temperature for 2 hours. The reaction was neutralized by theaddition of Amberlite IR 120 (H⁺) ion exchange resin. The solution wasfiltered through a glass fritted funnel with a pad of Celite to removethe resin. The filtrate was concentrated to dryness to give a crudetriol. The crude material was passed through a plug of silica (70:30 to85:15 EtOAc/hexanes) to give D-galactal triol (A126-2, 15.0 g 93%) aswhite solid. LC-MS (ESI) found: 147 [M+H]⁺.

Step 2: To a solution of(2R,3R,4R)-2-(hydroxymethyl)-3,4-dihydro-2H-pyran-3,4-diol (A126-2, 15.0g, 51.3 mmol) in DMF (200 mL) was added NaH (8.2 g, 205.3 mmol, 60% inmineral oil) at 0° C. in portions. The mixture was stirred at 0° C. for0.5 hours. Benzyl bromide (49.0 mL, 205.3 mmol) was added at 0° C. inportions. The mixture was stirred at room temperature for 2 hours. Themixture was quenched with H₂O, extracted with ethyl acetate, washed withH₂O, and concentrated and purified by column to give(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran(A126, 29.0 g, 68%) as white oil. LC-MS (ESI) found: 417 [M+H]⁺.

Synthesis 4-4. Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(Compound A127)

Step 1: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyranfrom Synthesis 4-3 (A126, 6.6 g, 15.8 mmol) and n-Bu₄NNO₃ (5.3 g, 17.4mmol) in dry DCM (70 mL) was added dropwise TFAA (3.7 g, 17.430 mmol) at0° C. under N₂ atmosphere. After the addition was complete, the reactionwas stirred at room temperature for 1 hour. Once the starting materialwas consumed (TLC monitoring), the reaction vessel was again cooled to0° C., TEA (2.4 mL, 17.4 mmol) was slowly added and the reaction mixturewas stirred for another 15 minutes. The reaction mixture was quenchedwith 10 mL ice water. Extraction was done with DCM (50 mL×3), and thecombined organic extracts were washed with water (50 mL×3) and brine(150 mL), dried over Na₂SO₄. The mixture was concentrated in vacuo. Thecrude product was purified by flash chromatography (silica gel, 0-20%EtOAc in PE) to give(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-nitro-3,4-dihydro-2H-pyran(A126-1, 4 g, 8.7 mmol, 54.7% yield) as a yellow oil. ¹H NMR (400 MHz,CDCl₃): δ 8.09 (s, 1H), 7.36-7.29 (m, 15H), 4.90 (dd, J=3.7, 1.3 Hz,1H), 4.86 (d, J=10.8 Hz, 1H), 4.79 (d, J=10.8 Hz, 1H), 4.70 (dd, J=13.4,8.5 Hz, 2H), 4.62 (d, J=11.9 Hz, 1H), 4.56 (d, J=11.9 Hz, 1H), 4.47 (d,J=11.9 Hz, 1H), 3.96-3.90 (m, 3H).

Step 2: To a solution of LiAlH₄ (0.66 g, 17.3 mmol) in dry THE (40 mL)was added(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-nitro-3,4-dihydro-2H-pyran(A126-1, 4 g, 8.7 mmol) in THE (10 mL) dropwise at 0° C. under N₂atmosphere. The reaction mixture was stirred at rt for 1 h. The reactionwas cooled to 0° C. and quenched with water (0.66 g), NaOH (0.66 g, 15%(w/w) in water), water (1.98 g). The mixture was filtered andconcentrated in vacuo. The crude product was purified by flashchromatography (silica gel, 0-90% EtOAc in PE) to give(4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A126-2, 1 g, 2.307 mmol, 26.6% yield) as a colorless oil, asapproximate 4:1 mixture of 3R/3S epimers. ¹H NMR (400 MHz, CDCl₃): δ7.39-7.27 (m, 15H), 4.90 (dd, J=22.9, 11.5 Hz, 1H), 4.74-4.68 (m, 1H),4.67-4.60 (m, 1H), 4.58-4.50 (m, 2H), 4.44 (d, J=11.9 Hz, 1H), 4.05 (dd,J=12.0, 2.0 Hz, 1H), 3.92-3.85 (m, 1H), 3.73-3.66 (m, 1H), 3.59-3.51 (m,3H), 3.48 (dd, J=8.4, 5.0 Hz, 1H), 3.15-3.07 (m, 1H).

Step 3: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A126-2. 7.4 g, 17.1 mmol) and DIPEA (8.5 mL, 51.2 mmol) in DCM (80 mL)was added acetyl chloride (2.4 mL, 34.1 mmol) dropwise at 0° C. Thereaction mixture was stirred at rt for 1.5 h. The resulting mixture wasdiluted with DCM (100 mL), washed with H₂O (80 mL×2) and brine (80 mL),dried over Na₂SO₄, filtered. The organic layer was separated andconcentrated in vacuo to give a crude product, which was purified byflash chromatography (silica gel, 0˜80% EA in PE) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(A127-3, 4.0 g, 8.4 mmol, 49.3%) as a colorless oil andN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(A127-4, 1.0 g, 2.1 mmol, 12.3% yield) as white solid.N-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(A127-3): LC-MS (ESI) found: 476 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃): δ7.38-7.28 (m, 15H), 7.20 (d, J=8.5 Hz, 1H), 4.88 (d, J=10.3 Hz, 1H),4.74 (d, J=11.8 Hz, 1H), 4.58 (d, J=11.8 Hz, 1H), 4.55-4.50 (m, 2H),4.48 (d, J=10.9 Hz, 2H), 4.01 (dd, J=12.2, 1.8 Hz, 1H), 3.95-3.92 (m,1H), 3.63 (d, J=6.4 Hz, 2H), 3.59 (dd, J=4.4, 2.9 Hz, 1H), 3.55-3.50 (m,1H), 3.46 (dd, J=12.2, 1.7 Hz, 1H), 1.77 (s, 3H).N-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(127-4): LC-MS (ESI) found: 476 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃): δ7.39-7.28 (m, 15H), 5.05 (s, 1H), 4.88 (d, J=11.5 Hz, 1H), 4.73 (d,J=12.2 Hz, 1H), 4.60 (d, J=11.6 Hz, 1H), 4.51 (d, J=11.9 Hz, 1H), 4.42(dd, J=14.7, 12.0 Hz, 2H), 4.24-4.16 (m, 2H), 4.00 (d, J=1.8 Hz, 1H),3.66-3.60 (m, 1H), 3.58-3.49 (m, 3H), 3.15 (t, J=11.9 Hz, 1H), 1.85 (s,3H).

Step 4: To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(A127-3, 100 mg, 0.2 mmol) in dry DCM (5 mL) was added BCl₃ (2.1 mL, 2.1mmol, 1 M in DCM) dropwise at −10° C. under N₂ atmosphere. The reactionmixture was stirred at rt for 0.5 h. Then the reaction was cooled to 0°C. and quenched with saturated sodium bicarbonate solution. The mixturewas filtered and concentrated in vacuo. The crude product was purifiedby flash chromatography (silica gel, 0-40% MeOH in DCM) to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(A127, 20 mg, 0.1 mmol, 46.4% yield) as a colorless oil. LC-MS (ESI)found: 206[M+H]⁺. ¹H NMR (400 MHz, CD3OD): δ4.08 (d, J=3.3 Hz, 1H),3.93-3.86 (m, 2H), 3.79-3.73 (m, 2H), 3.67 (dd, J=11.5, 4.8 Hz, 1H),3.53 (dd, J=12.0, 1.7 Hz, 1H), 3.44-3.37 (m, 1H).

Synthesis 4-5. Preparation ofN-((3aR,4R,7R,7aR)-4-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(Intermediate 5) andN-((4aR,7R,8R,8aR)-8-hydroxy-2,2-dimethylhexahydropyrano[3,2-d][1,3]dioxin-7-yl)acetamide(A127-2c)

To a solution ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamidefrom Synthesis 4-4 (A127, 100 mg, 0.49 mmol) in 2,2-dimethoxypropane (2mL) was added CSA (16.7 mg, 0.1 mmol) at room temperature under N₂. Thereaction was stirred at room temperature overnight. The crude productwas purified by prep-HPLC (Method A) to giveN-((3aR,4R,7R,7aR)-4-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(Intermediate 5, 19 mg, 16% yield) andN-((4aR,7R,8R,8aR)-8-hydroxy-2,2-dimethylhexahydropyrano[3,2-d][1,3]dioxin-7-yl)acetamide(A127-2c, 27 mg, 23% yield) as a colorless oil. LC-MS (ESI) of bothproducts found: 246[M+H]⁺.Synthesis 4-6. Preparation of(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A128)

To a solution ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamidefrom Synthesis 4-4 (A127, 20.0 mg, 0.09 mmol) in H₂O (2 mL) was addedBa(OH)₂ (166.0 mg, 0.97 mmol) at rt under N₂. The reaction mixture wasstirred at 100° C. overnight. The resulting mixture was filtered andconcentrated in vacuo. The crude product was purified by SCX cartridgesto give(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A128, 4.5 mg, 28%) as colorless oil. LC-MS (ESI) found: 164 [M+H]⁺. ¹HNMR (400 MHz, CD3OD): δ3.96 (dd, J=12.1, 1.9 Hz, 1H), 3.82-3.80 (m, 1H),3.76 (dd, J=11.4, 7.2 Hz, 1H), 3.68-3.64 (m, 1H), 3.62-3.58 (m, 2H),3.38-3.35 (m, 1H), 2.97 (dd, J=3.8, 1.8 Hz, 1H).Synthesis 4-7. Preparation of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(Compound A127-2a) and(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(Compound A127-2b)

75 mg of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A127-2a) and 20 mg of (3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A127-2b) were obtained by SFC separation from 100 mg of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A127-2 from Synthesis 4-4). The SFC was performed on Waters Thar 80preparative SFC (ChiralPak IC, 250×21.2 mm I.D., 5 μm; mobile phase: Afor CO₂ and B for MeOH+0.1% NH₃H₂O, gradient (40% B); Flow rate: 50mL/min, temperature: 35° C.).Synthesis 4-8. Preparation of tert-butyl((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)carbamate(Compound A129)

Step 1: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(A127-2a, 60 mg, 0.14 mmol) from Synthesis 4-7 in DCM (3 mL) was added(Boc)₂0 (60 mg, 0.28 mmol) at rt. After stirring at rt for 2 h, themixture was concentrated. Then the residue was purified by flashchromatography (silica gel, 0-20% EtOAc in PE) to give tert-butyl((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A129-1, 65 mg, 0.12 mmol, 88%) as a colorless oil. LC-MS (ESI) found:556 [M+Na]⁺. ¹H NMR (400 MHz, CDCl₃): δ 7.41-7.27 (m, 15H), 6.25 (d,J=9.1 Hz, 1H), 4.97 (d, J=10.7 Hz, 1H), 4.79 (d, J=11.8 Hz, 1H),4.50-4.46 (m, 4H), 4.20 (dd, J=9.0, 2.8 Hz, 1H), 4.04 (dd, J=12.1, 1.8Hz, 1H), 3.87 (t, J=7.0 Hz, 1H), 3.60-3.56 (m, 3H), 3.47 (dd, J=12.1,1.8 Hz, 1H), 1.57 (s, 9H).

Step 2: To a solution of tert-butyl((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A129-1, 60 mg, 0.11 mmol) in MeOH (2 mL) was added Pd/C (5 mg, 10% wt,60% wet). The reaction was charged with H₂ for three time and stirred atrt for 16 h. The mixture was filtered through a Celite pad, and thefiltrate was concentrated. The residue was purified by prep-HPLC (ColumnYMC Triart C18 250*20 mm, ID: 5 um. A: H₂O (0.1% FA), B: ACN, A % from95 to 55) to give tert-butyl((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)carbamate(A129, 20 mg, 0.076 mmol, 67.5%) as a colorless oil. LC-MS (ESI) found:264 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD): δ3.89 (dd, J=11.9, 1.4 Hz, 1H),3.85-3.82 (m, 1H), 3.75 (dd, J=11.4, 7.1 Hz, 2H), 3.72-3.69 (m, 1H),3.64 (dd, J=11.5, 4.8 Hz, 1H), 3.48 (dt, J=8.4, 4.2 Hz, 1H), 3.39-3.34(m, 1H), 1.44 (s, 9H).

Synthesis 4-9. Alternative Preparation of(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A128)

A solution of tert-butyl((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)carbamatefrom Synthesis 4-8 (A129, 10 mg, 0.04 mmol) in DCM(1 mL) was treatedwith HCl/dioxane (1 mL). The reaction mixture was stirred at rt for 3 hand then evaporated to give(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A130, 3.5 mg, 0.021 mmol, 56.5%) as a colorless oil. LC-MS (ESI) found:164 [M+H]⁺.

Synthesis 4-10. Preparation ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanesulfonamide(Compound A130)

To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(100 mg, 0.23 mmol) and TEA (0.1 mL, 0.69 mmol) in dry DCM (5 mL) at 0°C. under N₂ atmosphere was added dropwise MsCl (0.04 mL, 0.46 mmol). Thereaction mixture was stirred for 2 h. The resulting mixture was dilutedwith DCM (50 mL), washed with H₂O (20 mL×2) and brine (30 mL), driedover Na₂SO₄. The organic layer was separated and concentrated in vacuo.The crude product was purified by flash chromatography (silica gel,0˜80% EA in PE) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanesulfonamide(100 mg, 85% yield) as a colorless oil. LC-MS (ESI) found: 534 [M+Na]⁺.¹H NMR (400 MHz, CDCl₃): δ 7.40-7.27 (m, 15H), 6.09 (d, J=8.9 Hz, 1H),4.88 (d, J=11.0 Hz, 1H), 4.77 (d, J=11.7 Hz, 1H), 4.57 (dd, J=21.4, 11.7Hz, 3H), 4.45 (d, J=11.8 Hz, 1H), 4.06 (dd, J=12.2, 1.8 Hz, 1H), 3.95(dd, J=7.4, 2.3 Hz, 2H), 3.62 (dd, J=9.0, 6.0 Hz, 1H), 3.58-3.48 (m,4H), 2.89 (s, 3H).

Synthesis 4-11. Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methanesulfonamide(Compound A131)

To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanesulfonamide(50 mg, 0.1 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt, 60% wet)and HCl (1 mL, 1 M in H₂O) at rt under a H₂ balloon. The reaction wasstirred at rt for 3 h. The resulting mixture was filtered andconcentrated in vacuo. The crude product was purified by prep-HPLC(Method A) to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methanesulfonamide (1.9 mg, 8% yield) as a colorless oil. LC-MS (ESI)found: 242 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 3.99 (dd, J=12.1, 2.1 Hz,1H), 3.87-3.84 (m, 1H), 3.79-3.71 (m, 2H), 3.65 (dd, J=11.5, 4.8 Hz,1H), 3.60-3.54 (m, J=13.6, 2.1 Hz, 2H), 3.40-3.35 (m, J=6.9, 4.8, 1.3Hz, 1H), 3.04 (s, 3H).

Synthesis 4-12. Preparation ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(Compound A132)

Alternatively, Preparation ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(Compound A132) can be synthesized in the following manner:

To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(50 mg, 0.12 mmol) and TEA (0.05 mL, 0.35 mmol) in DCM (2 mL) was addedTf₂O (0.04 mL, 0.23 mmol) at 0° C. The reaction was stirred at rt for1.5 h. The resulting mixture was diluted with DCM (10 mL), washed withH₂O (5 mL×2) and brine (5 mL), dried over Na₂SO₄. The organic layer wasseparated and concentrated in vacuo. The crude product was purified byflash chromatography (silica gel, 0˜80% EA in PE) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(35 mg, 54% yield) as a colorless oil. LC-MS (ESI) found: 588 [M+Na]⁺.¹H NMR (400 MHz, CDCl₃): δ 7.40-7.26 (m, 15H), 7.00 (d, J=8.7 Hz, 1H),4.92 (d, J=10.7 Hz, 1H), 4.83 (d, J=11.9 Hz, 1H), 4.57-4.50 (m, 3H),4.44 (d, J=11.8 Hz, 1H), 4.06 (dd, J=12.5, 1.9 Hz, 1H), 4.00 (d, J=6.1Hz, 1H), 3.95-3.92 (m, 1H), 3.59 (dd, J=8.8, 5.8 Hz, 1H), 3.55-3.46 (m,4H).

Synthesis 4-13. Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(Compound A133)

Alternatively, Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(Compound A133) can be synthesized in the following manner:

To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(35 mg, 0.06 mmol) in MeOH (3 mL) was added Pd/C (4 mg, 10% wt, 60% wet)and HCl (1 mL, 1 M in H₂O) at rt under a H₂ balloon. The resultingmixture was filtered and concentrated in vacuo. The crude product waspurified by prep-HPLC (Method B) to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfonamide(10 mg, 55% yield) as a colorless oil. LC-MS (ESI) found: 294 [M−H]⁻. ¹HNMR (400 MHz, MeOD): δ 3.96 (dd, J=12.3, 2.1 Hz, 1H), 3.90-3.86 (m, 1H),3.80-3.73 (m, 2H), 3.72-3.63 (m, 2H), 3.60 (dd, J=12.3, 1.5 Hz, 1H),3.44-3.39 (m, 1H). ¹⁹F NMR (377 MHz, MeOD): δ-79.56 (s).

Synthesis 4-14. Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(Compound A134) andN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(Compound A135)

Step 1: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-aminefrom Synthesis 4-7 (A127-2, 1.0 g, 2.3 mmol) and TEA (0.8 mL, 5.8 mmol)in MeOH (5 mL) was added dropwise CF₃COOEt (0.41 mL, 3.46 mmol) at 0° C.and the reaction was 15 stirred at rt for 1.5 h. The resulting mixturewas diluted with DCM (50 mL), washed with H₂O (15 mL×2) and brine (20mL), dried over Na₂SO₄, filtered. The organic layer was concentrated invacuo to get a crude product, which was purified by flash chromatography(silica gel, 0-80% EA in PE) to give the mixture (1.0 g) as a colorlessoil. The mixture (100 mg) was then separated by SFC (OJ-H 4.6*250 mm,MeOH+0.05% DEA, 40%, 8 min) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A134-1, 50.0 mg) andN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A135-1, 15.0 mg).N-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A134-1): LC-MS (ESI) found: 530 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃): δ7.40-7.29 (m, 15H), 6.09 (d, J=5.9 Hz, 1H), 4.90 (d, J=11.5 Hz, 1H),4.71 (d, J=12.1 Hz, 1H), 4.62 (d, J=11.5 Hz, 1H), 4.54-4.44 (m, 2H),4.39 (d, J=12.1 Hz, 1H), 4.36-4.27 (m, 1H), 4.25-4.18 (m, 1H), 4.06 (d,J=2.2 Hz, 1H), 3.64-3.48 (m, 4H), 3.21 (t, J=10.8 Hz, 1H).N-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A-135-1): LC-MS (ESI) found: 530 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃): δ8.26 (d, J=8.3 Hz, 1H), 7.41-7.24 (m, 15H), 4.86 (d, J=10.6 Hz, 1H),4.72 (d, J=11.8 Hz, 1H), 4.57-4.44 (m, 5H), 4.04 (dd, J=12.5, 1.6 Hz,1H), 3.96 (s, 1H), 3.65-3.49 (m, 5H).

Step 2A: To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A134-1, 35 mg, 0.07 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedin vacuo. The crude product was purified by prep-HPLC (Method A) to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A134, 6.6 mg, 39%) as a white solid. LC-MS (ESI) found: 258 [M−H]⁻. ¹HNMR (400 MHz, CD3OD): δ4.28-4.15 (m, 1H), 3.95-3.86 (m, 2H), 3.76-3.61(m, 3H), 3.44-3.39 (m, 1H), 3.20 (t, J=11.0 Hz, 1H). ¹⁹F NMR (377 MHz,CD3OD): δ-77.23 (s).

Step 2B: To a solution ofN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A135-1, 15 mg, 0.03 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedin vacuo. The crude product was purified by (Method B) to giveN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A135, 4.9 mg, 66%) as a colorless oil. LC-MS (ESI) found: 258 [M−H]⁻.¹H NMR (400 MHz, MeOD): δ 4.12 (d, J=2.3 Hz, 1H), 3.97-3.89 (m, 2H),3.83-3.74 (m, 2H), 3.70-3.57 (m, 2H), 3.45-3.39 (m, 1H). ¹⁹F NMR (377MHz, MeOD): δ-78.03 (s).

Synthesis 4-15. Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(Compound A136) andN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(Compound A137)

Step 1: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-aminefrom Synthesis 4-7 (A127-2, 0.74 g, 1.7 mmol) and DIPEA (0.85 mL, 5.1mmol) in DCM (8.0 mL) was added benzoyl chloride (0.48 g, 3.4 mmol)dropwise at 0° C. The reaction mixture was stirred at rt for 1.5 h. Theresulting mixture was diluted with DCM (10 mL), washed with H₂O (10mL×2) and brine (10 mL), dried over Na₂SO₄, filtered. The organic layerwas concentrated in vacuo to give a crude product, which was purified byflash chromatography (silica gel, 0˜80% EA in PE) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(A136-1) andN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(A137-1). LC-MS (ESI) of both found: 538 [M+H]⁺.

Step 2A: To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(A136-1, 35 mg, 0.065 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedin vacuo.

The crude product was purified by prep-HPLC (Method A) to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(A136). LC-MS (ESI) of found: 268 [M+H]⁺.

Step 2B: To a solution ofN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(A137-1, 35 mg, 0.065 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedin vacuo. The crude product was purified by prep-HPLC (Method A) to giveN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(A136). LC-MS (ESI) of found: 268 [M+H]⁺.

Synthesis 4-16. Alternative Preparation of(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A128) and(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A92)

Step 1: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-aminefrom Synthesis 4-7 (A127-2, 0.74 g, 1.7 mmol) and DIPEA (0.85 mL, 5.1mmol) in DCM (8.0 mL) was added CbzCl (0.58 g, 3.4 mmol) dropwise at 0°C. The reaction mixture was stirred at rt for 1.5 h. The resultingmixture was diluted with DCM (10 mL), washed with H₂O (10 mL×2) andbrine (10 mL), dried over Na₂SO₄, filtered. The organic layer wasconcentrated in vacuo to give a crude product, which was purified byflash chromatography (silica gel, 0˜80% EA in PE) to give benzyl((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A128-1) and benzyl((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A92-4). LC-MS (ESI) of both found: 568 [M+H]⁺.

Step 2A: To a solution of benzyl((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A128-1, 35 mg, 0.06 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedto give(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A128). LC-MS (ESI) of both found: 164 [M+H]⁺.

Step 2B: To a solution of benzyl((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)carbamate(A92-4, 35 mg, 0.06 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedto give(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A92). LC-MS (ESI) of both found: 164 [M+H]⁺.

Synthesis 4-17. Preparation of(2R,3R,4R,5R)-5-((5-chloro-3-fluoropyridin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A138)

A solution of(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A128, 3.5 mg, 0.02 mmol), 5-chloro-2,3-difluoropyridine (8.9 mg, 0.06mmol) and DIPEA (11.6 mg, 15 0.09 mmol) in i-PrOH (4 mL) was stirred at120° C. overnight. The mixture was concentrated in vacuo. The crudeproduct was purified by pre-HPLC (Method A) to give(2R,3R,4R,5R)-5-((5-chloro-3-fluoropyridin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A138) as a white solid (1.0 mg, 16%). LC-MS (ESI) found: 293 [M+H]⁺.

The following compounds are prepared by the SNAr reaction in a methodsimilar to that of Synthesis 4-17 using either heating in isopropanol orNMP with Hunig's base and the corresponding commercially availablechloro or fluoro heterocycle.

Preparation of(2R,3R,4R,5R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl) tetrahydro-23-pyran-3,4-diol (Compound A146)

Step 1: To a solution of(2R,3R,4R,5R)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol (50mg, 0.3 mmol) and 3,5-dichloro-1,2,4-thiadiazole (142.5 mg, 0.92 mmol)in iPrOH (2 mL) was added DIPEA (0.25 mL, 1.53 mmol). The mixture wasstirred at 80° C. under N₂ overnight. The resulting mixture wasfiltered. The crude product was purified by flash chromatography (silicagel, 0˜10%0 MeOH in DCM) to give(2R,3R,4R,5R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol.(15 mg, 170 yield) as a colorless oil. LC-MS (ESI) found: 282 [M+H. ¹HNMR (400 MHz, MeOD): δ 4.08 (dd, J=12.2, 1.9 Hz, 1H), 4.01 (s, 1H),3.92-3.88 (m, 1H), 3.85-3.76 (m, 2H), 3.68 (dd, J=11.5, 4.7 Hz, 1H),3.60 (dd, J=12.2, 1.6 Hz, 1H), 3.46-3.42 (m, 1H).

ID Characterization data

Yield: 15 mg, 17%, colorless oil. LC-MS (ESI) found: 282 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.08 (dd, J = 12.2, 1.9 Hz, 1H), 4.01 (s, 1H),3.92-3.88 (m, 1H), 3.85-3.76 (m, 2H), 3.68 (dd, J = 11.5, 4.7 Hz, 1H),3.60 (dd, J = 12.2, 1.6 Hz, 1H), 3.46-3.42 (m, 1H).

Yield: 9.1 mg, 12.1%, colorless oil. LC-MS (ESI) found: 248 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 7.86 (s, 1H), 4.09 (dd, J = 12.2, 2.0 Hz, 1H),3.96 (s, 1H), 3.92-3.89 (m, 1H), 3.82 (ddd, J = 18.6, 8.0, 5.2 Hz, 2H),3.68 (dd, J = 11.5, 4.7 Hz, 1H), 3.59 (dd, J = 12.2, 1.6 Hz, 1H), 3.45(ddd, J = 6.9, 4.7, 1.2 Hz, 1H).

Yield: 5.1 mg, 5.9%, white solid. LC-MS (ESI) found: 281 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.35 (s, 1H), 4.08 (dd, J = 12.1, 2.0 Hz, 1H),3.92-3.89 (m, 2H), 3.81-3.76 (m, 2H), 3.67 (dd, J = 11.5, 4.7 Hz, 1H),3.54 (dd, J = 12.1, 1.6 Hz, 1H), 3.42 (ddd, J = 7.0, 4.7, 1.2 Hz, 1H).

Yield: 10 mg, 71%, colorless oil. LC-MS (ESI) found: 242 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.60 (s, 1H), 8.05 (s, 1H), 6.86 (d, J = 6.6 Hz,1H), 4.03 (d, J = 10.9 Hz, 1H), 3.94 (s, 1H), 3.91-3.88 (m, 1H), 3.82-3.65 (m, 4H), 3.50-3.45 (m, 1H).

Synthesis 4-18. Preparation of(2R,3R,4R,5R)-5-((3-(dimethylamino)-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A151)

Synthesis 4-19. General Synthesis of Amine-containing Compounds

Synthesis 4-20. Preparation ofN-((3S,4R,5R,6R)-6-(aminomethyl)-4,5-dihydroxy-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(Compound A53) andN-((3S,4R,5R,6R)-6-(aminomethyl)-4,5-dihydroxy-2-methoxytetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(Compound A154)

Alternatively,N-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(Compound A153) can be synthesized in the following manner:

Step 1: To a solution of(2R,3S,4R,5R,6R)-N-benzyl-4,5-bis(benzyloxy)-6-5((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine (280 mg, 0.506mmol) DMAP (12.36 mg, 0.101 mmol) and pyridine(0.818 mL, 10.120 mmol) inDCM (5 mL) was added acetic anhydride (0.475 mL, 5.060 mmol) atice-bath. The mixture was allowed to warm to rt and stirred for 3 h.Then the solvent was removed. The residue was purified by prep-HPLC(Method A) to giveN-benzyl-N-((2R,3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(160 mg, 53% yield). LC-MS (ESI) found: 596 [M+H]⁺.

Step 2: The solution ofN-benzyl-N-((2R,3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(100 mg, 0.198 mmol) in MeOH (5 mL) was added Pd/C (10 mg, 10% wt, 60%wet) and HCl (1 mL, 1 M in H₂O) at rt under a H₂ balloon. The mixturewas stirred overnight. The solvent was removed under vacuum.

The residue was purified by prep-HPLC (Method A) to give product (A153,5 mg, 10% yield) as colorless oil. LC-MS (ESI) found: 236 [M+H]⁺.

Example 5. Synthesis of ASGPR Ligands 20—Synthesis 5-1. GeneralSynthesis of Sulfonamide-containing-Ligands

Synthesis 5-2. General Synthesis of Sulfonyl Urea-Containing Compounds

Synthesis 5-3. Preparation ofN-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)carbamoyl)methanesulfonamide(Compound A157)

Step 1: To the solution of(3R,4R,5R,6R)-3-amino-6-(hydroxymethyl)tetrahydro-2H-pyran-2,4,5-triolhydrochloride (200 mg, 0.461 mmol) in THE (1 ml) and H₂O (9 ml) wasdropwise added diphenyl carbonate (119 mg, 0.554 mmol). The mixture wasstirred at rt overnight. The mixture was extracted with EA, washed withNaOH solution, dried over Na₂SO₄, concentrated and purified by silica(silica gel, 0-100% EA in PE) to give phenylN-[2,4,5-trihydroxy-6-(hydroxymethyl)oxan-3-yl] carbamate (100 mg, 39%yield) as solid. LC-MS (ESI) found: 554 [M+H]⁺.

Step 2: A solution of phenylN-[4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxan-3-yl]carbamate (20 mg,0.036 mmol), methanesulfonamide (7 mg, 0.072 mmol) and DBU (17 mg, 0.108mmol) in CH₃CN (5 mL) was stirred at 80° C. overnight. The mixture wasconcentrated and purified by column (silica gel, 0-10% MeOH in DCM) togive1—[4,5-bis(benzyloxy)-6-[(benzyloxy)methyl]oxan-3-yl]—3-methanesulfonylurea(10 mg, 50% yield) as white solid. LC-MS (ESI) found: 555 [M+H]⁺.

Step 3: To the solution of1—[4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxan-3-yl]—3-methanesulfonylurea(10 mg, 0.018 mmol) in MeOH (2 mL) was added Pd/C (4 mg, 10% wt, 60%wet), the mixture was stirred at rt overnight under a H₂ balloon. Themixture was filtered and purified by prep-HPLC (Method A) to giveN-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)carbamoyl)methanesulfonamide(9 mg, 46%) as white solid. LC-MS (ESI) found: 285 [M+H]⁺. ¹H NMR (400MHz, MeOD): δ 4.03 (dd, J=10.6, 5.2 Hz, 1H), 3.95 (dd, J=10.5, 5.0 Hz,1H), 3.86 (d, J=2.7 Hz, 1H), 3.72 (dd, J=10.7, 7.7 Hz, 1H), 3.65 (dd,J=11.3, 4.9 Hz, 1H), 3.50 (dd, J=10.3, 3.1 Hz, 1H), 3.44-3.38 (m, 1H),3.22 (s, 1H), 3.12 (t, J=10.5 Hz, 1H).

Synthesis 5-4. General Synthesis of Sulfonimidamide-containing Ligands

Synthesis 5-5. Alternative General Synthesis ofSulfonimidamide-containing Ligands

Synthesis 5-6. General Synthesis of ASGPR Ligands

Synthesis 5-7. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-phenethyltetrahydro-2H-pyran-3,4-diol(Compound A161)

Step 1: solution of 2R,3R-3,4- bis(benzyloxy)-2- (benzyloxy)methyl]-3,4-dihydro-2H-pyran (A126, 6.0 g, 14.4 mmol), NIS (3.8 g, 17.3 mmol) andAgNO₃ (0.5 g, 2.9 mmol) in CH₃CN (100 mL) was stirred at 80° C. for 30min. The mixture was filtered and concentrated to give a yellow solid,which was purified by column chromatography on silica gel to give(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(A122-1, 3.4 g, 44%) as a white solid. LC-MS (ESI) found: 543 [M+H]⁺. HNMR (400 MHz, CD3OD): δ7.45-7.22 (m, 15H), 6.63 (d, J=1.2 Hz, 1H),4.81-4.67 (m, 3H), 4.60-4.38 (m, 3H), 4.34 (t, J=7.0 Hz, 1H), 4.20 (t,J=5.5 Hz, 1H), 4.13 (dt, J=12.2, 6.1 Hz, 1H), 3.76-3.60 (m, 2H).

Step 2: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(A122-1, 400 mg, 0.74 mmol) in THE (3 mL) was added CuI (14 mg, 0.074mmol), TEA (223.45 mg, 2.212 mmol), Pd(PPh₃)₂Cl₂ (85 mg, 0.074 mmol) andethynylbenzene (0.12 mL, 1.106 mmol). The mixture was charged with N₂for three times and stirred at 60° C. under N₂ atmosphere overnight. Themixture was concentrated and purified by flash eluting with PE/EA (from95/5 to 50/50) to give desired product (A160-1, 360 mg, 95%) asoff-white solid. LC-MS (ESI) found: 517 [M+H]⁺.

Step 3: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(phenylethynyl)-3,4-dihydro-2H-pyran(160-1, 25 mg, 0.048 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet). The mixture was stirred at rt under H₂ atmosphere for 12 h.The mixture was filtered and concentrated to give 4.1 mg of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-phenethyltetrahydro-2H-pyran-3,4-diol(A160). LC-MS (ESI) found: 253 [M+H]⁺. ¹H NMR (400 MHz, CD3OD):δ7.55-6.74 (m, 5H), 3.95-3.84 (m, 2H), 3.83-3.77 (m, 1H), 3.74 (t, J=3.1Hz, 1H), 3.66 (dd, J=11.8, 4.0 Hz, 1H), 3.51 (dt, J=7.3, 3.6 Hz, 1H),3.43 (dd, J=11.7, 3.0 Hz, 1H), 2.70 (ddd, J=13.6, 10.3, 5.8 Hz, 1H),2.54 (ddd, J=13.6, 9.8, 6.4 Hz, 1H), 2.05-1.76 (m, 2H), 1.70 (dt, J=9.2,4.5 Hz, 1H). nOe experiment was consistent with the indicatedstereochemistry of the product.

Using the procedure shown in Synthesis 5-7,(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(3-hydroxypropyl)tetrahydro-2H-pyran-3,4-diol(Compound A162) andN-(3-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)propyl)acetamide(Compound A163) were prepared

Synthesis 5-8. Preparation of(2R,3R,4R,5R)-5-(4-fluorophenyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A164):

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyranfrom Synthesis 5-7 (A122-1, 100 mg, 0.184 mmol) in DME (10 mL) was addedPd(PPh₃)₄(21 mg, 0.018 mmol), K₂CO₃ (76 mg, 0.552 mmol) and(4-fluorophenyl)boronic acid (34 mg, 0.239 mmol). The mixture wascharged with N₂ for three times and stirred at 90° C. under N₂ for 16 h.The mixture was filtered, the filtrate was concentrated under reducedpressure to give a crude product, which was purified by flashchromatography (silica gel, 5-10% EtOAc in PE) to give(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(4-fluorophenyl)-3,4-dihydro-2H-pyran(A164-1, 25 mg, 31% yield) as colorless oil. LC-MS (ESI) found: 533[M+23]⁺.

Step 2: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(4-fluorophenyl)-3,4-dihydro-2H-pyran(A164-1, 20 mg, 0.039 mmol) in MeOH (5 mL) was added Pd/C (5 mg, 10% wt,60% wet). The mixture was stirred at rt for 12 h under H₂ atmosphere.The mixture was filtered and concentrated to give(2R,3R,4R,5R)-5-(4-fluorophenyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A164, 5.0 mg, 53% yield). LC-MS (ESI) found: 243 [M+H]⁺. ¹H NMR (400MHz, CD3OD): δ7.56-7.42 (m, 2H), 7.04-6.88 (m, 2H), 4.23 (dd, J=11.7,7.1 Hz, 1H), 4.09 (td, J=9.0, 2.8 Hz, 1H), 4.01-3.95 (m, 1H), 3.92-3.85(m, 1H), 3.76 (ddd, J=13.7, 7.4, 3.6 Hz, 2H), 3.71-3.62 (m, 1H),3.02-2.91 (m, 1H).

Using the procedures shown in Synthesis 5-8,(2R,3R,4R,5R)-5-(2,4-difluorophenyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A165),(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1H-pyrazol-3-yl)tetrahydro-2H-pyran-3,4-diol(Compound A166) and(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1H-pyrazol-4-yl)tetrahydro-2H-pyran-3,4-diol(Compound A167) were prepared

Synthesis 5-9. Alternative General Synthesis of ASGPR Ligands:

Synthesis 5-6 and Synthesis 5-9 can be used to synthesize ligands withthe following R² groups:

wherein R is an optimal substituent has defined herein.Synthesis 5-10: Preparation of(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-N,N-dimethyltetrahydro-2H-pyran-3-carboxamide(Compound A170) and(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxylicacid (Compound A169)

Step 1: A solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanol(A170-1, 500 mg, 1.115 mmol), NaIO₄ (978 mg, 4.570 mmol) and RuCl₃(0.002 mL, 0.028 mmol) in CCl₄ (10 mL), H₂O (15 mL), and MeCN (10 mL)was stirred at rt for 2 h. The mixture was extracted with DCM (10 mL×3).The combined organic phase was washed with brine, dried over Na₂SO₄,filtered and concentrated in vacuo to give a crude product, which waspurified by column to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (A169-1, 400 mg, 78%) as white solid. LC-MS (ESI) found: 463[M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 7.35-7.19 (m, 15H), 4.75-4.62 (m, 3H),4.59-4.39 (m, 3H), 4.28-4.22 (m, 1H), 4.05 (dd, J=12.0, 7.8 Hz, 1H),3.93 (ddd, J=22.1, 13.1, 5.9 Hz, 2H), 3.81 (dd, J=4.0, 2.6 Hz, 1H),3.66-3.57 (m, 2H), 2.84 (dt, J=8.0, 4.2 Hz, 1H).

Step 2: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (A169-1, 60 mg, 0.13 mmol) in THE (5 mL) was added EDCI (30 mg,0.16 mmol), HOBT (26 mg, 0.19 mmol), and dimethylamine (0.13 mmol, 1 Min THF). The mixture was stirred at rt for 12 h. The mixture wasextracted by DCM (10 mL×3). The combined organic phase was washed withbrine, dried over Na₂SO₄, filtered and concentrated in vacuo to give acrude product, which was purified by silica gel column to give desiredproduct (A170-2). LC-MS (ESI) found: 490 [M+H]⁺.

Step 3A: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (A169-1, 30 mg, 0.065 mmol) in MeOH (3 mL) was added Pd/C (5 mg,10% wt, 60% wet). The mixture was stirred at rt for 12 h under H₂atmosphere. The mixture was filtered and concentrated to give(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxylicacid (A169), 3.0 mg. LC-MS (ESI) found: 193 [M+H]⁺.

Alternatively, Step 3A can be performed in the following manner: To asolution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (15 mg, 0.032 mmol) in MeOH (1 mL) was added Pd/C (10 mg, 10% wt,60% wet). The mixture was stirred at rt for 12 h under a H₂ balloon. Themixture was filtered and concentrated to give(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxylicacid (6 mg, 96%) as a yellow solid. LC-MS (ESI) found: 193 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.27 (d, J=10.8 Hz, 1H), 3.96 (dd, J=5.7, 3.5 Hz,1H), 3.84-3.72 (m, 2H), 3.72-3.59 (m, 2H), 3.46-3.36 (m, 1H), 2.86 (d,J=20.2 Hz, 1H).

Step 3B: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N,N-dimethyltetrahydro-2H-pyran-3-carboxamide(A170-2, 30 mg, 0.061 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt,60% wet). The mixture was stirred at rt for 12 h under H₂ atmosphere.The mixture was filtered and concentrated to give 2 mg of(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-N,N-dimethyltetrahydro-2H-pyran-3-carboxamide(A170). LC-MS (ESI) found: 220 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 4.04(dd, J=12.3, 0.8 Hz, 1H), 3.97 (dt, J=15.6, 7.8 Hz, 1H), 3.76 (dt,J=13.4, 6.7 Hz, 1H), 3.69 (ddd, J=12.4, 6.4, 4.2 Hz, 3H), 3.47 (dd,J=5.9, 3.6 Hz, 1H), 3.39-3.32 (m, 1H), 3.14 (s, 3H), 2.99 (s, 3H).

Using the procedure shown in Synthesis 5-10,1-(4-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carbonyl)piperazin-1-yl)ethan-1-one(Compound A171),((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)(4-methylpiperazin-1-yl)methanone(Compound A172),((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)(piperidin-1-yl)methanone(Compound A173), and(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxamide(Compound A174) were prepared

Alternatively,(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxamide(Compound A174) can be synthesized in the following manner:

Step 1: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (A169-1, 100 mg, 0.22 mmol) in DMF (10 mL) was added HATU (107 mg,0.28 mmol). After stirring at rt for 1 h, NH₄Cl (23 mg, 0.43 mmol) andTEA (65 mg, 0.65 mmol) were added. The mixture was stirred at rt for 1h, then diluted with ethyl acetate and washed with water. The organiclayer was dried over sodium sulfate, filtered, concentrated underreduced pressure to give a crude product, which was purified by silicagel column to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxamide(20 mg, 20% yield). LC-MS (ESI) found: 462 [M+H]⁺.

Step 2: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxamide(20 mg, 0.043 mmol) in MeOH (10 mL) was added Pd/C (20 mg, 10% wt, 60%wet). The mixture was stirred at rt overnight under a H₂ balloon. Themixture was filtered, the filtrate was concentrated to give(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxamide(3.5mg, 42% yield). LC-MS (ESI) found: 192 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ3.92 (dd, J=11.4, 4.8 Hz, 1H), 3.77-3.67 (m, 2H), 3.59 (ddd, J=16.4,11.4, 6.0 Hz, 2H), 3.39-3.26 (m, 2H), 2.72 (m, 1H).

Synthesis 5-11. Preparation of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N′-(2,2,2-trifluoroacetyl)tetrahydro-2H-pyran-3-carbohydrazide(Compound A175)

Alternatively,(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N′—(2,2,2-trifluoroacetyl)tetrahydro-2H-pyran-3-carbohydrazide(Compound A175) can be synthesized in the following manner:

Step 1: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxane-3-carboxylicacid(400 mg, 0.865 mmol) and CDI (0.12 mL, 0.951 mmol) in DCM (20 mL)was added hydrazine hydrate (4.2 mL, 86.5 mmol) at 0° C. The mixture wasstirred at rt for 12 h. the mixture was concentrated and purified bycolumn to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carbohydrazide(140 mg, 34% yield) as yellow solid. LC-MS (ESI) found: 477 [M+H]⁺.

Step 2: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxane-3-carbohydrazide(140 mg, 0.30 mmol) and TFAA (0.08 mL, 0.60 mmol) in MeCN (10 mL) wasadded DIPEA (46 mg, 0.353 mmol) at 0° C. The mixture was stirred at rtfor 2 h. The mixture was concentrated and purified by column to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N′—(2,2,2-trifluoroacetyl)tetrahydro-2H-pyran-3-carbohydrazide(135 mg, 80% yield) as yellow oil. LC-MS (ESI) found: 573 [M+H]⁺.

Synthesis 5-12. Preparation of2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-5-(trifluoromethyl)-1,3,4-oxadiazole(Compound A176)

Synthesis 5-13. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(5-(trifluoromethyl)-1,3,4-oxadiazol-2-yl)tetrahydro-2H-pyran-3,4-diol(Compound A177)

Synthesis 5-14. Preparation of(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-N-(methylsulfonyl)tetrahydro-2H-pyran-3-carboxamide(Compound A178)

Synthesis 5-15. General Synthesis of Amide-Containing Ligands

Synthesis 5-16: Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyrrolidin-2-one(Compound A180)

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(A122-1, 100 mg, 0.184 mmol) in DMF (3 mL) was added CuI (3.4 mg, 0.018mmol), Cs₂CO₃ (120 mg, 0.368 mmol), pyrrolidin-2-one (43 mg, 0.552mmol). The mixture was stirred at 160° C. under microwave for 1 h. Themixture was filtered, the filtrate was concentrated under reducedpressure to give a crude product, which was purified by flashchromatography to give1-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)pyrrolidin-2-one(A180-1, 27 mg, 30% yield) as colorless oil. LC-MS (ESI) found: 500[M+1]⁺.

Step 2: To a solution of1-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)pyrrolidin-2-one(A180-1, 20 mg, 0.04 mmol) in MeOH (5 mL) was added Pd/C (5 mg, 10% wt,60% wet). The mixture was stirred at rt for 12 h under H₂ atmosphere.The mixture was filtered and concentrated to give 3 mg of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyrrolidin-2-one(A180). LC-MS (ESI) found: 232 [M+H]⁺.

Alternatively,1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyrrolidin-2-one(Compound A180) can be synthesized in the following manner:

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(100 mg, 0.18 mmol) in DMF (30 mL) was added pyrrolidin-2-one (78 mg,0.92 mmol), Cs₂CO₃ (180 mg, 0.55 mmol), CuI (4 mg, 0.021 mmol), themixture was stirred at 160° C. under microwave for 1 h. The mixture wasdiluted with ethyl acetate and washed with water. The organic layer wasdried over sodium sulfate, filtered, concentrated under reduced pressureto give a crude product, which was purified by silica gel column to give1-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)pyrrolidin-2-one(20 mg, 21% yield). LC-MS (ESI) found: 500 [M+H]⁺.

Step 2: To a solution of1-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)pyrrolidin-2-one(20 mg, 0.04 mmol) in MeOH (10 mL) was added Pd/C (20 mg, 10% wt, 60%wet), the mixture was stirred at rt under a H₂ balloon overnight. Themixture was filtered, the filtrate was concentrated to give1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyrrolidin-2-one(2.3 mg, 25% yield). LC-MS (ESI) found: 232 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 4.18-4.06 (m, 2H), 3.97 (dd, J=12.7, 3.0 Hz, 1H), 3.77 (m, 2H),3.70-3.66 (m, 1H), 3.59 (m, 2H), 3.46-3.35 (m, 2H), 2.32-2.23 (m, 2H),1.90 (m, 2H).

Synthesis 5-17: Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-methyltetrahydro-2H-pyran-3,4-diol(Compound A181) and(2R,3R,4R,5R)-2,5-bis(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A182)

Step 1: To a solution of DMF (50 mL) was added POCl₃ (6.2 mL, 66.3 mmol)at 0° C. in portions. The mixture was stirred at 0° C. for 0.5 h.(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyranfrom Synthesis 4-3 (A126, 9.2 g, 22.1 mmol) in DMF (20 mL) was added at0° C. in portions. The mixture was stirred at rt for 5 h. The mixturewas quenched with H₂O, extracted with EA, washed with H₂O and brine, andconcentrated and purified by silica gel column to give(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-carbaldehyde(A182-1, 6.0 g, 61%) as yellow oil. LC-MS (ESI) found: 445 [M+H]⁺. ¹HNMR (400 MHz, CD3OD): δ9.29 (s, 1H), 7.40-7.14 (m, 15H), 4.76-4.64 (m,4H), 4.62-4.54 (m, 2H), 4.48 (q, J=12.0 Hz, 2H), 4.00-3.80 (m, 3H).

Step 2: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-carbaldehyde(A182-1, 10.0 g, 22.4 mmol) in MeOH (50 mL) was added NaBH₄ (17.0 g,45.0 mmol) at 0° C. in portions. The mixture was stirred at rt for 12 h.The mixture was quenched with H₂O, extracted with EA, and concentratedto give crude((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanol(A182-2, 9.5 g, 95% yield) as yellow oil. LC-MS (ESI) found: 469[M+Na]⁺. ¹H NMR (400 MHz, CD3OD); 6 7.38-7.21 (m, 15H), 6.37 (s, 1H),4.78 (t, J=10.9 Hz, 2H), 4.63 (dd, J=20.6, 11.4 Hz, 2H), 4.54-4.37 (m,3H), 4.28-4.14 (m, 2H), 4.08-4.02 (m, 1H), 3.89 (d, J=11.9 Hz, 1H),3.75-3.65 (m, 2H).

Step 3: To a solution of((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanol(A182-2, 6.0 g, 13.4 mmol) in MeOH (2 mL) was added Pd/C (1.2 g, 10% wt,60% wet) in portions. The mixture was stirred at rt under H₂ for 2 h.The mixture was filtered, concentrated, and purified by silica column togive((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanol(A170-1, 0.49 g, 8% yield), LC-MS (ESI) found: 449 [M+H]⁺, and(2R,3R,4R,5R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methyltetrahydro-2H-pyran(A182-3, 1.1 g, 19% yield) as colorless oil, LC-MS (ESI) found: 433[M+H]⁺.

Step 4A: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanol(A170-1, 60.0 mg, 0.134 mmol) in MeOH(2 mL) was added Pd/C (30 mg, 10%wt, 60% wet). The mixture was stirred at rt under H₂ for 12 h. Themixture was filtered and concentrated to give(2R,3R,4R,5R)-2,5-bis(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol (A181,6.3 mg, 26%) as brown oil. LC-MS (ESI) found: 179 [M+H]⁺. ¹H NMR (400MHz, CD3OD): δ4.08 (dd, J=11.8, 2.0 Hz, 1H), 3.89 (ddd, J=9.1, 8.4, 5.5Hz, 2H), 3.81 (dd, J=11.0, 4.0 Hz, 1H), 3.75 (dt, J=11.3, 5.6 Hz, 2H),3.64 (dd, J=11.5, 4.6 Hz, 1H), 3.50 (dd, J=11.8, 2.7 Hz, 1H), 3.37 (ddd,J=13.0, 7.4, 5.1 Hz, 1H), 1.88 (d, J=2.9 Hz, 1H).

Step 4B: To a solution of(2R,3R,4R,5R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methyltetrahydro-2H-pyran(A182-3, 50 mg, 0.11 mmol) in MeOH (2 mL) was added Pd/C (10 mg, 10% wt,60% wet). The mixture was stirred at rt under H₂ for 2 h. Then thereaction mixture was filtered and concentrated to give(2R,3R,4R)-2,5-bis(hydroxymethyl)oxane-3,4-diol (A182, 8 mg, 40%) as awhite solid. LC-MS (ESI) found: 185 [M+Na]⁺. ¹H NMR (400 MHz, CD3OD):δ3.85 (dd, J=11.7, 7.6 Hz, 1H), 3.79-3.62 (m, 4H), 3.55-3.42 (m, 2H),1.93-1.79 (m, 1H), 1.11 (d, J=7.2 Hz, 3H).

Synthesis 5-18. Preparation of((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl4-methylbenzenesulfonate (Compound A183)

Step 1: To a solution of((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanolfrom Synthesis 5-17 (A170-1, 350 mg, 0.78 mmol) and TEA (394 mg, 3.90mmol) in DCM (10 mL) was added TsCl (446 mg, 2.34 mmol) slowly at 0° C.The reaction mixture was stirred at rt overnight. The resulting mixturewas extracted with EA (50 mL), washed with H₂O (40 mL×2) and brine (40mL), dried over Na₂SO₄, filtered. The organic layer was separated andconcentrated in vacuo. The crude product was purified by flashchromatography (silica gel, 0˜50% EA in PE) to give((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl4-methylbenzenesulfonate (A183-1, 180 mg, 38%) as a yellow oil. LC-MS(ESI) found: 603 [M+H]⁺.

Step 2: To a solution of(2R,3R,4R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyran(A183-1, 185 mg, 0.31 mmol) in dry DMF (10 mL) was added NaN₃ (180 mg,3.10 mmol). The reaction mixture was stirred at 80° C. overnight. Themixture was extracted with EA (20 mL), washed with H₂O (20 mL×2) andbrine (20 mL), dried over Na₂SO₄, filtered. The organic layer wasseparated and concentrated in vacuo. The crude product was purified byflash chromatography (silica gel, 0˜50% EA in PE) to give(2R,3R,4R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyran(A183-2, 65 mg, 45% yield) as a colorless oil. LC-MS (ESI) found:496[M+Na]⁺.

Step 3: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(A183-3, 65 mg, 0.14 mmol) in THE (5 mL) was added PPh₃ (72 mg, 0.27mmol) and water (5 mL). The reaction mixture was stirred at rtovernight. The mixture was concentrated in vacuo. The crude product waspurified by flash chromatography (silica gel, 0-10% MeOH in DCM) to give((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(A183-3, 60 mg, 98% yield) as a colorless oil. LC-MS (ESI) found:448[M+H]⁺.

Step 4: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(A183-3, 30 mg, 0.067 mmol) in dry DCM (5 mL) was added DIPEA (30 mg,0.07 mmol) and acetyl chloride (11 mg, 0.14 mmol) dropwise at 0° C.under N₂ atmosphere. The reaction mixture was stirred at rt for 8 h. Thereaction was cooled to 0° C. and quenched with saturated sodiumbicarbonate solution. The mixture was extracted with EA (10 mL), washedwith H₂O (10 mL×2) and brine (10 mL), dried over Na₂SO₄, filtered. Theorganic layer was separated and concentrated in vacuo. The crude productwas purified by flash chromatography (silica gel, 0˜50% EA in PE) togiveN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(A183-4, 14 mg, 44% yield) as a colorless oil. LC-MS (ESI) found:490[M+H]⁺.

Step 5: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(A183-4, 14 mg, 0.03 mmol) in dry MeOH (5 mL) was added Pd/C (10 mg, 10%wt, 60% wet). The reaction mixture was charged with H₂ and stirred at rtfor 3 days under H₂ atmosphere. The mixture was filtered andconcentrated in vacuo to giveN-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(A183, 1 mg, 16% yield) as a colorless oil. LC-MS (ESI) found:220[M+H]⁺. H NMR (400 MHz, CD3OD): δ3.91 (dd, J=11.9, 2.8 Hz, 1H),3.85-3.72 (m, 3H), 3.66 (dd, J=11.7, 4.3 Hz, 1H), 3.58 (dd, J=13.9, 4.3Hz, 1H), 3.48-3.40 (m, 3H), 1.93 (s, 3H), 1.92-1.84 (m, 1H).

Synthesis 5-19. Preparation of(2R,3R,4R)-5-(azidomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A184)

To a solution of(2R,3R,4R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyranfrom Synthesis 5-18 (A183-2, 20 mg, 0.04 mmol) in dry DCM (5 mL) at −78°C. under N₂ atmosphere was added BCl₃ (0.4 mL, 0.04 mmol, 1 M in DCM)slowly. After the addition was complete, the reaction was stirred at 0°C. for 45 min. On consumption of starting material (TLC monitoring), thereaction mixture was quenched with 1 mL MeOH. The mixture wasconcentrated in vacuo. The crude product was purified by pre-HPLC(Method A) to give(2R,3R,4R)-5-(azidomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A184, 0.7 mg, 8% yield) as a colorless oil. LC-MS (ESI) found: 226[M+Na]⁺. ¹H NMR (400 MHz, CD3OD): δ3.99 (dd, J=11.9, 2.0 Hz, 1H), 3.82(dd, J=5.4, 3.3 Hz, 1H), 3.79-3.74 (m, 2H), 3.73-3.67 (m, 2H), 3.64 (dd,J=11.5, 4.5 Hz, 1H), 3.48-3.44 (m, 1H), 3.40-3.36 (m, 1H), 1.97-1.84 (m,1H).

Synthesis 5-20. Preparation of(2R,3R,4R,5R)-5-(aminomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A185)

To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanaminefrom Synthesis 5-18 (A183-3, 20 mg, 0.044 mmol) in dry MeOH (5 mL) wasadded Pd/C (5 mg, 10% wt, 60% wet). The reaction mixture was chargedwith H₂ and stirred at rt for 3 days under H₂ atmosphere. Then themixture was filtered and concentrated in vacuo to give(2R,3R,4R,5R)-5-(aminomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A185) as a colorless oil. LC-MS (ESI) found: 178 [M+H]⁺.Alternatively,(2R,3R,4R,5R)-5-(aminomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A185) can be synthesized in the following manner:

Step 1: To a solution of(2R,3R,4R,5R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-5((benzyloxy)methyl)tetrahydro-2H-pyran (A183-2, 50 mg, 0.11 mmol) inMeOH (5 mL) were added Pd/C (5 mg, 10% wt, 60% wet) and HCl (0.1 mL, 1 Nin H₂O). The reaction mixture was stirred at rt overnight under aballoon of H₂. The mixture was filtered and concentrated in vacuo togive(2R,3R,4R,5R)-5-(aminomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A185, 16 mg, 86% yield) as yellow oil. LC-MS (ESI) found: 178 [M+H]⁺.¹H NMR (400 MHz, 10 MeOD): δ4.09-4.02 (m, 1H), 3.96 (dd, J=5.7, 3.3 Hz,1H), 3.85 (d, J=2.7 Hz, 1H), 3.76-3.70 (m, 1H), 3.69-3.61 (m, 2H),3.43-3.33 (m, 2H), 3.26 (dd, J=13.0, 3.7 Hz, 1H), 2.22-2.05 (m, 1H).

Synthesis 5-21. Preparation ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-2,2,2-trifluoroacetamide(Compound A186)

Step 1: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanaminefrom Synthesis 5-18 (A183-3, 30 mg, 0.067 mmol) in dry MeOH (5 mL) wasadded ethyl 2,2,2-trifluoroacetate (19 mg, 0.134 mmol) and triethylamine(26 mg, 0.201 mmol) at rt. The reaction mixture was stirred at rtovernight. The resulting mixture was diluted with EA (10 mL), washedwith H₂O (10 mL×2) and brine (10 mL), dried over Na₂SO₄, filtered. Theorganic layer was concentrated in vacuo to get a crude product, whichwas purified by flash chromatography (silica gel, 0˜50% EA in PE) togiveN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-2,2,2-trifluoroacetamide(A186-1, 16 mg, 44%) as a yellow oil. LC-MS (ESI) found: 566 [M+Na]⁺.

Step 2: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-2,2,2-trifluoroacetamide(A186-1, 16 mg, 0.029 mmol) in dry MeOH (5 mL) was added Pd/C (3 mg, 10%wt, 60% wet). The reaction mixture was charged with H₂ for three timesand stirred at rt for 3 days under H₂ atmosphere. The mixture wasfiltered and concentrated in vacuo to give 3 mg ofN-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)-2,2,2-trifluoroacetamide(A186) as colorless oil. LC-MS (ESI) found: 274/296 [M+H]⁺/[M+Na]⁺.

Alternatively, Step 2 can be performed in the following manner: To asolution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-2,2,2-trifluoroacetamide(16 mg, 0.029 mmol) in MeOH (5 mL), Pd/C (5 mg, 10% wt, 60% wet) and HCl(0.1 mL, 1 N in H₂O) was added. The reaction mixture was stirred at rtovernight under a H₂ balloon. The mixture was filtered, the filtrate wasconcentrated to giveN-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)-2,2,2-trifluoroacetamide(A186, 7.9 mg, 99%) as yellow oil. LC-MS (ESI) found: 274 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 3.91 (dd, J=12.1, 1.9 Hz, 1H), 3.86 (dd, J=5.6, 3.3Hz, 1H), 3.81-3.79 (m, 1H), 3.78-3.74 (m, 1H), 3.73-3.68 (m, 1H),3.67-3.61 (m, 2H), 3.51 (dd, J=12.1, 2.8 Hz, 1H), 3.39 (ddd, J=7.1, 4.5,1.7 Hz, 1H), 2.03-1.96 (m, 1H).

Synthesis 5-22. Preparation ofN-((1-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)acetamide(Compound A187)

Step 1: A solution of THPTA (0.32 mg, 0.001 mmol) and CuSO₄ (2.4 mg,0.02 mmol) in H₂O (0.5 mL) was added to a solution of(2R,3R,4R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyranfrom Synthesis 5-18 (A183-2, 32.0 mg, 0.07 mmol) andN-(prop-2-yn-1-yl)acetamide (8.6 mg, 0.09 mmol) in MeOH (2 mL). Afreshly-prepared solution of sodium ascorbate (5.9 mg, 0.03 mmol) in H₂O(0.5 mL) was added and the reaction mixture was stirred at roomtemperature for 24 h. Then the mixture was concentrated and the residualwas purified by flash chromatography (0-5% methanol in dichloromethane)to giveN-((1-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)acetamide(A187-1, 33 mg, 78%) as oil. LC-MS (ESI) found: 571 [M+H]⁺.

Step 2: To a solution ofN-((1-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)acetamide(A187-1, 16 mg, 0.03 mmol) in MeOH (3 mL) was added Pd/C (2 mg, 10% wt,60% wet), the mixture was charged with H₂ for three times and stirred atthe room temperature under H₂ atmosphere overnight. The mixture wasfiltered, the filtrate was concentrated under reduced pressure to give acrude product, which was purified by flash chromatography (0-50% ethylacetate in petroleum ether) to giveN-((1-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1H-1,2,3-triazol-4-yl)methyl)acetamide(A187, 2 mg, 24%). LC-MS (ESI) found: 301 [M+H]⁺. ¹H NMR (400 MHz,CD3OD): δ7.85 (s, 1H), 4.83 (s, 1H), 4.68 (dd, J=14.1, 1.9 Hz, 1H), 4.42(s, 2H), 3.91 (dd, J=5.3, 3.2 Hz, 1H), 3.82 (dd, J=11.5, 7.1 Hz, 2H),3.75-3.68 (m, 2H), 3.49-3.40 (m, 2H), 2.33-2.23 (m, 1H), 1.98 (s, 3H).

Synthesis 5-23. Preparation of(2R,3R,4R,5R)-5-(((3-chloro-1,2,4-thiadiazol-5-yl)amino)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A188)

Step 1: A solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanaminefrom Synthesis 5-18 (A183-3, 50 mg, 0.112 mmol),3,5-dichloro-1,2,4-thiadiazole (51 mg, 0.336 mmol) and DIPEA (44 mg,0.336 mmol) in i-PrOH (5 mL) was stirred at rt overnight. The mixturewas concentrated in vacuo. The crude product was purified by pre-HPLC(Method A) to giveN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-3-chloro-1,2,4-thiadiazol-5-amine(A188-1, 25 mg, 40%). LC-MS (ESI) found: 566 [M+H]⁺

Step 2: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-3-chloro-1,2,4-thiadiazol-5-amine(A188-1, 25 mg, 0.044 mmol) in dry DCM (5 mL) was added BCl₃ (0.44 mL, 1M in DCM) dropwise at −10° C. under N₂ atmosphere. The reaction mixturewas stirred at rt for 0.5 h. Then the reaction was cooled to 0° C. andquenched with saturated sodium bicarbonate solution. The mixture wasfiltered and concentrated in vacuo. The crude product was purified byprep-HPLC (Method A) to give 7 mg of(2R,3R,4R,5R)-5-(((3-chloro-1,2,4-thiadiazol-5-yl)amino)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A188). LC-MS (ESI) found: 296 [M+H]⁺.

Alternatively,(2R,3R,4R,5R)-5-(((3-chloro-1,2,4-thiadiazol-5-yl)amino)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A188) can be synthesized in the following manner:

Step 1: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(30 mg, 0.067 mmol) in i-PrOH (5 mL) were added3,5-dichloro-1,2,4-thiadiazole (33 mg, 0.21 mmol) and DIPEA (43 mg, 0.34mmol). The reaction mixture was stirred at rt overnight. The resultingmixture was extracted with EA (10 mL), washed with H₂O and brine, driedover Na₂SO₄. The organic layer was separated and concentrated in vacuo.The crude product was purified by flash chromatography to giveN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-3-chloro-1,2,4-thiadiazol-5-amine(10 mg, 27%) as yellow oil. LC-MS (ESI) found: 566 [M+H]⁺.

Step 2: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-3-chloro-1,2,4-thiadiazol-5-amine(10 mg, 0.018 mmol) in dry DCM (5 mL), BCl₃ (0.2 mL, 1 N in DCM) wasadded under the −78° C. The reaction mixture was stirred at 0° C. for 45min. The resulting mixture was extracted with DCM (10 mL), washed withH₂O and brine, dried over Na₂SO₄. The crude product was purified byPre-TLC to give(2R,3R,4R,5R)-5-(((3-chloro-1,2,4-thiadiazol-5-yl)amino)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(5 mg, 67%) as yellow oil. LC-MS (ESI) found: 296 [M+H]⁺. ¹H NMR (400MHz, MeOD): δ 3.97 (dd, J=12.1, 2.0 Hz, 1H), 3.87 (dd, J=5.4, 3.3 Hz,1H), 3.83-3.77 (m, 2H), 3.76-3.55 (m, 3H), 3.50 (dd, J=12.1, 1.9 Hz,1H), 3.44-3.39 (m, 1H), 2.09-2.02 (m, 1H).

Synthesis 5-24. Preparation of(E)-2-((((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)amino)ethene-1-sulfonylfluoride (Compound A189)

Synthesis 5-25. Preparation of2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetonitrile(Compound A190)

Step 1: To a solution of((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl4-methylbenzenesulfonate from Synthesis 5-18 (A183-1, 200 mg, 0.46 mmol)in DMSO (5 mL) was added NaCN (68 mg, 1.39 mmol) at room temperature.The mixture was stirred at rt overnight. Then the mixture was extractedby DCM (10 mL * 3) and the organic phase was concentrated. The residualwas then purified by silica gel column (0-22% ethyl acetate in petroleumether) to give2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetonitrile(A190-1, 80 mg, 38%) as an oil. LC-MS (ESI) found: 458 [M+H]⁺.

Step 2: To a solution of2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetonitrile(A190-1. 15 mg, 0.03 mmol) in dry DCM (3 mL) was added BCl₃ (0.33 mL,0.33 mmol, 1 M in DCM) dropwise at −10° C. under N₂ atmosphere. Thereaction mixture was stirred at rt for 0.5 h. Then the reaction wascooled to 0° C. and quenched with saturated sodium bicarbonate solution.The mixture was filtered and concentrated in vacuo. The crude productwas purified by flash chromatography (silica gel, 0-40% MeOH in DCM) togive2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetonitrile(A190, 2 mg, 33%) as a colorless oil. LC-MS (ESI) found: 188 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.05 (dd, J=12.3, 1.8 Hz, 1H), 3.80 (ddd, J=16.3,8.4, 4.5 Hz, 3H), 3.68 (d, J=4.3 Hz, 1H), 3.66-3.58 (m, 2H), 3.42 (ddd,J=7.2, 4.5, 1.6 Hz, 1H), 2.98 (dd, J=17.4, 11.3 Hz, 1H), 2.85 (ddd,J=17.4, 3.8, 1.3 Hz, 1H).

Synthesis 5-26. Preparation of(2R,3R,4R,5S,6S)-2-(hydroxymethyl)-5-methyl-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A191),(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-5-methyl-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A192), and(2R,3R,4R,5S)-2-(hydroxymethyl)-5-(phenoxymethyl)tetrahydro-2H-pyran-3,4-diol

Step 1: To a solution of((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanolfrom Synthesis 5-17 (A182-2, 100 mg, 0.224 mmol, 1.0 eq), PPh₃ (88 mg,0.336 mmol, 1.5 eq) and nucleophiles (1.0 eq) in dry DCM (1.1 mL) wasadded DIAD (0.053 mL, 0.269 mmol, 1.2 eq) dropwise at ice-bath under N₂atmosphere. Then the reaction was allowed to warm to rt. The resultingreaction mixture was stirred at the same temperature for another 40 min,at which time TLC showed the disappearance of all starting material. Themixture was evaporated. The crude product was further purified by silicagel column chromatography to give desired products. Yield (A192-1): 30mg, 26%. LC-MS (ESI) found: 545 [M+Na]⁺. Yield (A193-1): 70 mg, 60%.LC-MS (ESI) found: 545 [M+Na]⁺.

Step 2A: A suspension of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methylene-6-phenoxytetrahydro-2H-pyran(A192-1, 100 mg, 1.0 eq) and Pd/C (0.2 eq, 10% wt, 60% wet) in MeOH wascharged with H₂ and stirred under H₂ atmosphere. The reaction wasstirred at rt and monitored by TLC. When TLC showed the disappearance ofall starting material, the mixture was filtered and evaporated. Thecrude product was further purified by silica gel column chromatographyto give desired products. Yield (A192): 0.5 mg, 2%. Yield (A191): 7 mg,14%. LC-MS (ESI) found: 277 [M+Na]⁺. ¹H NMR (400 MHz, CD3OD) 6 7.30-7.22(m, 2H), 7.13-7.06 (m, 2H), 6.96 (tt, J=7.4, 1.1 Hz, 1H), 5.44 (d, J=1.8Hz, 1H), 4.11 (dd, J=5.5, 3.5 Hz, 1H), 3.89 (d, J=2.2 Hz, 2H), 3.75-3.68(m, 2H), 2.20 (ddd, J=7.4, 5.3, 1.8 Hz, 1H), 1.23 (d, J=7.4 Hz, 3H).

Step 2B:(2R,3R,4R,5S)-2-(hydroxymethyl)-5-(phenoxymethyl)tetrahydro-2H-pyran-3,4-diol)(A193) was synthesized according to the hydrogenation proceduredescribed for Step 2A above. Yield: 9.8 mg, 20%. LC-MS (ESI) found: 277[M+Na]⁺. ¹H NMR (400 MHz, Methanol-d₄): δ 7.30-7.19 (m, 2H), 6.95-6.84(m, 3H), 4.37 (t, J=10.0 Hz, 1H), 4.24 (ddd, J=9.7, 3.0, 1.4 Hz, 1H),4.14 (dd, J=11.7, 1.9 Hz, 1H), 3.91 (dd, J=5.6, 3.2 Hz, 1H), 3.81-3.74(m, 2H), 3.66 (dd, J=11.5, 4.6 Hz, 1H), 3.51 (ddd, J=11.7, 2.6, 1.4 Hz,1H), 3.41 (ddd, J=7.2, 4.6, 1.7 Hz, 1H), 2.22 (ddd, J=10.2, 5.4, 2.7 Hz,1H).

Synthesis 5-27. Preparation of(2R,3R,4R,5S,6S)-2-(hydroxymethyl)-5-methyl-6-(1H-tetrazol-1-yl)tetrahydro-2H-pyran-3,4-diol(Compound A194),(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-5-methyl-6-(1H-tetrazol-1-yl)tetrahydro-2H-pyran-3,4-diol(Compound A195), and(2R,3R,4R,5R)-5-((1H-tetrazol-1-yl)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A196)

Step 1:1-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-3-methylenetetrahydro-2H-pyran-2-yl)-1H-tetrazole(A195-1) and1-(((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methyl)-1H-tetrazole(A196-1) were synthesized according to the Mitsunobu procedure describedin Synthesis 5-26 using 500 mg of((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanol(A182-2) from Synthesis 5-17.1-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-3-methylenetetrahydro-2H-pyran-2-yl)-1H-tetrazole(A195-1) Yield: 50 mg, 9%. LC-MS (ESI) found: 521 [M+Na]⁺.1-(((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methyl)-1H-tetrazole(A196-1) Yield: 15 mg, 5%. LC-MS (ESI) found: 521 [M+Na]⁺.

Step 2A:(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-5-methyl-6-(1H-tetrazol-1-yl)tetrahydro-2H-pyran-3,4-diol(A195) and(2R,3R,4R,5S,6S)-2-(hydroxymethyl)-5-methyl-6-(1H-tetrazol-1-yl)tetrahydro-2H-pyran-3,4-diol(A194) were synthesized according to the hydrogenation proceduredescribed in Synthesis 5-26 using 65 mg of1-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-3-methylenetetrahydro-2H-pyran-2-yl)-1H-tetrazole(A195-1): Yield((2R,3R,4R,5S,6R)-2-(hydroxymethyl)-5-methyl-6-(1H-tetrazol-1-yl)tetrahydro-2H-pyran-3,4-diol)(A195): 3 mg, 10%. LC-MS (ESI) found: 253 [M+Na]⁺. Yield((2R,3R,4R,5S,6S)-2-(hydroxymethyl)-5-methyl-6-(1H-tetrazol-1-yl)tetrahydro-2H-pyran-3,4-diol)(A194): 1 mg, 3%. LC-MS (ESI) found: 253 [M+Na]⁺.

Step 2B:(2R,3R,4R,5R)-5-((1H-tetrazol-1-yl)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A196) was synthesized according to the hydrogenation proceduredescribed in Synthesis 5-26 using 14.4 mg of1-(((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methyl)-1H-tetrazole(A196-1; Yield((2R,3R,4R,5R)-5-((1H-tetrazol-1-yl)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol)(A196): 2 mg, 30%. LC-MS (ESI) found: 231 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 8.68 (s, 1H), 5.20 (dd, J=14.0, 11.0 Hz, 1H), 5.05-4.93 (m,1H), 3.93 (dd, J=5.5, 3.2 Hz, 1H), 3.87-3.76 (m, 3H), 3.70 (dd, J=11.5,4.6 Hz, 1H), 3.50-3.38 (m, 2H), 2.54-2.22 (m, 1H).

Synthesis 5-28.(2R,3R,4R,5R)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)-1H-pyrazol-1-yl)methyl)tetrahydro-2H-pyran-3,4-diol(Compound A197),1-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)-1H-pyrazole-4-carbonitrile(Compound A198),(2R,3R,4R,5R)-5-((4-chloro-1H-pyrazol-1-yl)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A199), and(2R,3R,4R,5R)-5-((1H-pyrazolo[3,4-b]pyridin-1-yl)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A200) were prepared using the procedure shown in Synthesis5-26

Synthesis 5-29. Preparation of(2R,3R,4R,5S,6S)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A200) and(2R,3R,4R,5S,6R)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A201)

Step 1: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methylene-6-phenoxytetrahydro-2H-pyranfrom Synthesis 5-26 (A192-1, 100 mg, 0.19 mmol) in DCM (10 mL) was addedprop-2-en-1-ol (110 mg, 1.9 mmol) and Grubbs catalyst 2^(nd) generation(16 mg, 0.019 mmol). The reaction was charged with N₂ for three time andstirred at 40° C. for 16 h. LCMS showed the desired product wasdetected. The solvent was concentrated in vacuo to get a crude product,which was purified by column to afford(Z)-2-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-phenoxydihydro-2H-pyran-3(4H)-ylidene)ethan-1-ol(A200-1) as yellow oil. LC-MS (ESI) found: 553 [M+H]⁺. Reference:Journal of the American Chemical Society, 123(42), 10417-10418; 2001.

Step 2: A solution of(Z)-2-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-phenoxydihydro-2H-pyran-3(4H)-ylidene)ethan-1-ol(A200-1, 50 mg, 0.09 mmol) and Pd/C (5 mg, 10% wt, 60% wet) in MeOH wasstirred under H₂ atmosphere. The reaction was stirred at r.t overnightThen the mixture was filtered and evaporated. The crude product wasfurther purified by silica gel column chromatography to give(2R,3R,4R,5S,6S)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(A200) and(2R,3R,4R,5S,6R)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(A201). LC-MS (ESI): 285 [M+H]⁺.

Synthesis 5-30. Preparation of(2R,3R,4R,5S,6S)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A202) and(2R,3R,4R,5S,6R)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A203)

Step 1: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methylene-6-phenoxytetrahydro-2H-pyranfrom Synthesis 5-26 (A192-1, 100 mg, 0.19 mmol) in DCM (10 mL was addedN-allylacetamide (188 mg, 1.9 mmol) and Grubbs catalyst 2^(nd)generation (16 mg, 0.019 mmol). The reaction was charged with N₂ forthree time and stirred at 40° C. for 16 h. LCMS showed the desiredproduct was detected. The solvent was concentrated in vacuo to get acrude product, which was purified by column to affordN-((Z)-2-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-phenoxydihydro-2H-pyran-3(4H)-ylidene)ethyl)acetamide(A202-1). LC-MS (ESI) found: 594 [M+H]⁺. Reference: Journal of theAmerican Chemical Society, 123(42), 10417-10418; 2001.

Step 2: A solution of(Z)-2-((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-phenoxydihydro-2H-pyran-3(4H)-ylidene)ethan-1-ol(A202-1, 50 mg, 0.08 mmol) and Pd/C (5 mg, 10% wt, 60% wet) in MeOH wasstirred under H₂ atmosphere. The reaction was stirred at r.t overnightThen the mixture was filtered and evaporated. The crude product wasfurther purified by silica gel column chromatography to give(2R,3R,4R,5S,6S)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A202) and(2R,3R,4R,5S,6R)-5-(2-hydroxyethyl)-2-(hydroxymethyl)-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A203). LC-MS (ESI): 326 [M+H]⁺.

Synthesis 5-31. Preparation of di-tert-butyl2-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)malonate(Compound A204)

Synthesis 5-32. Preparation of3-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)propanoicacid (Compound A205)

Synthesis 5.33. Preparation ofN-(2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)ethyl)acetamide(Compound A206) andN-(2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)ethyl)-2,2,2-trifluoroacetamide(Compound A207)

Step 1: A solution of2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetonitrile(A190-1, 100 mg, 0.22 mmol) and Raney Ni (15 mg) in MeOH was chargedwith H₂ for three times and stirred under H₂ atmosphere overnight Thenthe mixture was filtered and evaporated. The crude product was furtherpurified by silica gel column chromatography to give2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)ethan-1-amine(A206-1). LC-MS (ESI): 462 [M+H]⁺.

Step 2: To a solution of2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)ethan-1-amine(A206-1, 80 mg, 0.17 mmol) in MeOH (5 mL) was added Pd/C (10 mg, 10% wt,60% wet). The reaction was charged with H₂ for three time and stirredunder H₂ atmosphere overnight. The crude product was obtained byfiltration and concentration. LC-MS (ESI) found for both targets: 192[M+H]⁺.

Step 3A: To a solution of(2R,3R,4R,5R)-5-(2-aminoethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A206-2, 15 mg, 0.078 mmol) in THF (2 mL) was added AcC1 (9.1 mg, 0.118mmol) and TEA (23.6 mg, 0.234 mmol) at rt. The mixture was stirred at rtfor 2 h. The mixture was concentrated and the crude material waspurified by silica gel column to giveN-(2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)ethyl)acetamide(A206). LC-MS (ESI) found: 234 [M+H]⁺.

Step 3B: To a solution of(2R,3R,4R,5R)-5-(2-aminoethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A206-2, 15 mg, 0.078 mmol) in MeOH (2 mL) was added CF₃COOEt (16.7 mg,0.118 mmol) and TEA (23.6 mg, 0.234 mmol) at rt. The mixture was stirredat rt for 2 h. The mixture was concentrated and the crude material waspurified by silica gel column to giveN-(2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)ethyl)-2,2,2-trifluoroacetamide(A207). LC-MS (ESI) found: 288 [M+H]⁺.

Synthesis 5-34. Preparation of3-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)-1,1-dimethylurea(Compound A208)

Step 1: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanaminefrom Synthesis 5-18 (A183-3, 100 mg, 0.224 mmol) in DCM (5 mL) was added4-nitrophenyl carbonochloridate (67.4 mg, 0.336 mmol) and DIPEA (87.0mg, 0.672 mmol) at rt. The mixture was stirred at rt overnight. Themixture was concentrated and used directly for next step. LC-MS (ESI)found: 613 [M+H]⁺.

Step 2: To a solution of 4-nitrophenyl(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)carbamate(A208-1, 137 mg, 0.224 mmol) in DCM (5 mL) was added dimethylamine(0.448 mmol, 1 M in THF) and DIPEA (87.0 mg, 0.672 mmol) at rt. Themixture was stirred at rt overnight. The mixture was concentrated andpurified by silica gel column to give 30 mg of the desired product(A208-2). LC-MS (ESI) found: 519 [M+H]⁺.

Step 3: To a solution of3-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1,1-dimethylurea(A208-2, 30 mg, 0.058 mmol) in MeOH (3 mL) was added Pd/C (6 mg, 10% wt,60% wet). The reaction was charged with H₂ for three times and stirredunder H₂ atmosphere overnight. The crude product was obtained byfiltration and concentration. LC-MS (ESI): 249 [M+H]⁺.

Alternatively,3-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)-1,1-dimethylurea(Compound A208) can be synthesized in the following manner:

Step 1: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(45 mg, 0.110 mmol) in dry DCM (10 mL), 4-nitrophenyl carbonochloridate(34 mg, 0.170 mmol) and DIPEA (28 mg, 0.220 mmol) was added. Thereaction mixture was stirred at rt overnight. The resulting mixture wasextracted with EA (10 mL), washed with H₂O (10 mL×2) and brine (10 mL),dried over Na₂SO₄ to give crude 4-nitrophenyl(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)carbamate(67 mg) as a yellow oil. LC-MS (ESI) found: 613 [M+H]⁺.

Step 2: To a solution of 4-nitrophenyl(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)carbamate(67 mg, 0.110 mmol) in dry DCM (5 mL), dimethylamine (30 mg, 0.667 mmol)and DIPEA (60 mg, 0.467 mmol) was added. The reaction mixture wasstirred at rt overnight. The resulting mixture was extracted with EA (10mL), washed with H₂O (10 mL×2) and brine (10 mL), dried over Na₂SO₄. Theorganic layer was separated and concentrated in vacuo. The crude productwas purified by flash chromatography (silica gel, 0˜50% EA in PE) togive3-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1,1-dimethylurea(25 mg, 44% yield) as a yellow oil. LC-MS (ESI) found: 519 [M+H]⁺.

Step 3: To a solution of3-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)-1,1-dimethylurea(25 mg, 0.048 mmol) in dry MeOH (5 mL), Pd/C (5 mg, 10% wt, 60% wet) andHCl (0.1 mL, 1 M in H₂O) was added. The reaction mixture was stirred atrt overnight under a H₂ balloon. The organic mixture was filtration andconcentrated in vacuo to give3-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)-1,1-dimethylurea(9 mg, 76% yield) as a yellow oil. LC-MS (ESI) found: 249 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 3.93 (dd, J=12.0, 2.1 Hz, 1H), 3.91-3.86 (m, 1H),3.85-3.76 (m, 2H), 3.70-3.61 (m, 2H), 3.55-3.40 (m, 3H), 2.95 (s, 6H),2.01-1.92 (m, 1H).

Synthesis 5-35. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(morpholinomethyl)tetrahydro-2H-pyran-3,4-diol(Compound A209)

Step 1: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanolfrom Synthesis 5-17 (A192-2, 500.0 mg, 1.11 mmol) in DCM (5 mL) wasadded DMP (945.5 mg, 2.22 mmol) at 0° C. in portions. The mixture wasstirred at rt for 5 h. The mixture was quenched with aqueous NaHCO₃. Thetwo phases were separated and the organic phase was dried over Na₂SO₄,filtered and concentrated to give a crude product, which was purified bycolumn to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carbaldehyde(A209-1, 350.2 mg, 70%) as yellow oil. LC-MS (ESI) found: 447 [M+H]⁺. ¹HNMR (400 MHz, CD3OD): δ9.93 (s, 1H), 7.47-7.14 (m, 15H), 4.81-4.37 (m,8H), 4.17-3.95 (m, 2H), 3.79-3.62 (m, 1H), 3.59-3.41 (m, 2H), 2.70 (s,1H).

Step 2: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxane-3-carbaldehyde(A209-2, 60.0 mg, 0.134 mmol) and morpholine(0.024 mL, 0.269 mmol) inDCM (2 mL) was added NaBH(OAc)₃ (170 mg, 0.269 mmol) at 0° C. inportions. The mixture was stirred at rt for 2 h. The mixture wasquenched with H₂O and concentrated in vacuo to give a crude product,which was purified by prep-HPLC to give4-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)morpholine(A209-2, 62 mg, 89%) as colorless oil. LC-MS (ESI) found: 518 [M+H]⁺.

Step 3: To a solution of4-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)morpholine(A290-2, 60.0 mg, 0.116 mmol) in MeOH (2 mL) was added Pd/C (30 mg, 10%wt, 60% wet). The mixture was stirred at rt for 12 h under H₂atmosphere. The mixture was filtered and concentrated to give(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(morpholinomethyl)tetrahydro-2H-pyran-3,4-diol(A209, 4.2 mg, 15%) as yellow oil. LC-MS (ESI) found: 248 [M+H]⁺. ¹H NMR(400 MHz, MeOD); 6 3.87-3.81 (m, 2H), 3.76 (dd, J=6.8, 4.7 Hz, 2H), 3.68(ddd, J=13.9, 9.3, 4.4 Hz, 6H), 3.56 (dd, J=12.1, 3.2 Hz, 1H), 3.37(ddd, J=7.2, 4.5, 1.5 Hz, 1H), 3.23 (dd, J=12.8, 9.4 Hz, 1H), 2.68-2.52(m, 4H), 2.14 (d, J=4.5 Hz, 1H).

Synthesis 5-36. Preparation of1-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-N,N-dimethylmethanamine(Compound A2010)

Step 1: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxane-3-carbaldehydefrom Synthesis 5-35 (A209-1, 60 mg, 0.134 mmol) and dimethylaminehydrochloride (33 mg, 0.403 mmol) in DCM (2 mL) was added NaBH(OAc)₃(170 mg, 0.134 mmol) at 0° C. in portions. The mixture was stirred at rtfor 2 h. The mixture was quenched with H₂O and concentrated to give acrude product, which was purified by prep-HPLC (Method A) to give1-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-N,N-dimethylmethanamine(A210-1, 60 mg, 94%) as colorless oil. LC-MS (ESI) found: 476 [M+H]⁺. ¹HNMR (400 MHz, CD3OD): δ7.42-7.20 (m, 15H), 4.79 (d, J=11.2 Hz, 1H), 4.66(d, J=12.4 Hz, 2H), 4.56-4.41 (m, 3H), 3.94 (dd, J=11.9, 2.7 Hz, 1H),3.89-3.80 (m, 2H), 3.70-3.61 (m, 2H), 3.52 (ddd, J=13.9, 12.7, 6.0 Hz,2H), 2.98-2.86 (m, 1H), 2.76 (s, 1H), 2.29 (s, 6H), 2.16 (d, J=7.2 Hz,1H).

Step 2: To a solution of1-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-N,N-dimethylmethanamine(A210-1, 60 mg, 0.126 mmol) in MeOH (3 mL) was added Pd/C (10 mg, 10%wt, 60% wet). The mixture was stirred at rt for 12 h under H₂atmosphere. The mixture was filtered and concentrated to give(2R,3R,4R,5R)-5-((dimethylamino)methyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A210) as yellow oil. LC-MS (ESI) found: 206 [M+H]⁺.

Synthesis 5-37.1-(4-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)piperazin-1-yl)ethan-1-one(Compound A211),(2R,3R,4R,5R)-2-(hydroxymethyl)-5-((4-methylpiperazin-1-yl)methyl)tetrahydro-2H-pyran-3,4-diol(Compound A212), and(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(piperidin-1-ylmethyl)tetrahydro-2H-pyran-3,4-diol(Compound A213) were prepared using the procedure of Synthesis 5-36.

A211

A212

A213

ID Characterization data A212 LC-MS (ESI) found: 261 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 4.06-3.54 (m, 16H), 3.46-3.41 (m, 1H), 3.03 (s, 3H),2.51-2.42 (m, 1H). A213 LC-MS (ESI) found: 246 [M + H]⁺. ¹H NMR (400MHz, MeOD): δ 3.99 (dd, J = 11.4, 4.6 Hz, 1H), 3.80 (d, J = 2.6 Hz, 1H),3.73-3.65 (m, 3H), 3.60 (dd, J = 10.6, 3.1Hz, 2H), 3.42-3.37 (m, 1H),3.23-3.18 (m, 1H), 3.18-3.14 (m, 1H), 3.05-2.93 (m, 3H), 2.50-2.42 (m,1H), 1.99-1.93 (m, 2H), 1.86-1.75 (m, 3H), 1.59-1.52 (m, 1H).Synthesis 5-38. Preparation of(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A214)

Step 1: To a mixture of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyranfrom Synthesis 4-3 (A126, 6.5 g, 15.6 mmol) and1-dodecyl-3-methylImidazolium tetrafluoroborate (1.06 g, 3.1 mmol) inDCM (65 mL) was added acetone (26 mL) and NaHCO₃ (49 mL). Then oxone(19.5 g, mmol) in H₂O (81 mL) was added dropwise to the stirringreaction at 0° C. After 2 hours, the mixture was quenched with H₂O andextracted with EA. The combined organic layer was washed with brine,dried over anhydrous Na₂SO₄, filtered. The filtrate was concentrated togive(1S,3R,4S,5S,6R)-4,5-bis(benzyloxy)-3-((benzyloxy)methyl)-2,7-dioxabicyclo[4.1.0]heptane(A214-1, 6.0 g, 89%) as a colorless oil. LC-MS (ESI) found: 433 [M+H]⁺.

Step 2: A solution of(1S,3R,4S,5S,6R)-4,5-bis(benzyloxy)-3-((benzyloxy)methyl)-2,7-dioxabicyclo[4.1.0]heptane(A214-1, 6.0 g, 13.9 mmol) in MeOH (0.6 mL, 13.9 mmol) was stirred at rtovernight. The mixture was concentrated and purified by chromatographyon (silical gel, 0-50% ethyl acetate in petroleum ether) to give(2R,3R,4R,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-ol(A214-2, 4.5 g, 70%) as a white solid. LC-MS (ESI) found: 465 [M+H]⁺.

Step 3: To a mixture of(2R,3R,4R,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-ol(A214-2, 200 mg, 0.43 mmol) in dry DCM (5.0 mL) was added dry pyridine(0.53 mL) and Tf₂O (705 mg, 0.43 mmol) at 0° C. under N₂. After stirringfor 2 h, the mixture was concentrated and purified by chromatography on(0-30% ethyl acetate in petroleum) to give(2R,3R,4S,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yltrifluoromethanesulfonate (A214-3, 137 mg, 53.3%) as a yellow oil. LC-MS(ESI) found: 597 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 7.37-7.25 (m, 15H),5.23 (dd, J=9.8, 3.7 Hz, 1H), 4.96 (d, J=3.7 Hz, 1H), 4.89 (d, J=11.2Hz, 1H), 4.74-4.62 (m, 2H), 4.51-4.39 (m, 3H), 4.05-3.98 (m, 2H), 3.92(t, J=6.5 Hz, 1H), 3.53 (t, J=5.2 Hz, 2H), 3.43 (d, J=10.1 Hz, 3H).

Step 4: A solution of(2R,3R,4S,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yltrifluoromethanesulfonate (A214-3, 200 mg, 0.336 mmol) and benzyl amine(360 mg, 3.36 mmol) in dry THE (2 mL) was heated to 80° C. in seal tube.The resulting reaction mixture was monitored by TLC. When TLC showed thedisappearance of all starting material, the mixture was evaporated. Thecrude product was further purified by silica gel column chromatography(PE/EA=1:1) to give desired product (A214-4, 250 mg, 67% yield) asyellow oil. LC-MS (ESI) found: 554[M+H]⁺.

Step 5: To a solution of(2R,3S,4R,5R,6R)-N-benzyl-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A214-4, 35 mg, 0.063 mmol) in MeOH (3 mL) was added Pd/C (10 mg, 10%wt, 60% wet) at rt under H₂. The reaction was stirred at rt under H₂atmosphere for 3 h. The resulting mixture was filtered and concentratedin vacuo to give(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A214) as a white solid. LC-MS (ESI) found: 194[M+H]⁺.

Synthesis 5-39. Preparation ofN-((2R,3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)acetamide(Compound A215)

Synthesis 5-40. Alternative Preparation of(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A214)

Step 1:(2R,3S,4R,5R,6R)-3-azido-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran(A215-2) was prepared following a similar procedure for(2R,3S,4R,5R,6R)-N-benzyl-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A214-4) in Synthesis 5-38. Yield: 300 mg, 73%. LC-MS (ESI) found: 512[M+Na]⁺.

Synthesis 5-41: Preparation of(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-6-methoxy-5-(methylamino)tetrahydro-2H-pyran-3,4-diol(Compound A216)

Step 1:(2R,3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxy-N-methyltetrahydro-2H-pyran-3-amine(A216-1) was prepared following a similar procedure for(2R,3S,4R,5R,6R)-N-benzyl-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A214-4) in Synthesis 5-38. LC-MS (ESI) found: 478 [M+H]⁺.

Step 2:(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-6-methoxy-5-(methylamino)tetrahydro-2H-pyran-3,4-diol(A216) was prepared following a similar procedure for(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A214) in Synthesis 5-38. LC-MS (ESI) found: 208 [M+H]⁺.

Synthesis 5-42. Preparation of(2R,3R,4S,5S,6R)-2-(hydroxymethyl)-5,6-dimethoxytetrahydro-2H-pyran-3,4-diol(Compound A217)

Step 1:(2R,3S,4S,5S,6R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5,6-dimethoxytetrahydro-2H-pyran(A217-1) was prepared following a similar procedure for(2R,3S,4R,5R,6R)-N-benzyl-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A214-4) in Synthesis 5-38. Yield: 170 mg, 83%. LC-MS (ESI) found: 501[M+Na]⁺.

Step 2:(2R,3R,4S,5S,6R)-2-(hydroxymethyl)-5,6-dimethoxytetrahydro-2H-pyran-3,4-diol(A217) was prepared following a similar procedure for(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A214) in Synthesis 5-38. Yield: 15 mg, 55%. LC-MS (ESI) found: 231[M+Na]⁺. ¹H NMR (400 MHz, Methanol-d₄) δ 4.33 (d, J=4.6 Hz, 1H), 4.17(p, J=5.0 Hz, 2H), 4.03-3.97 (m, 1H), 3.89 (t, J=4.9 Hz, 1H), 3.79-3.68(m, 2H), 3.44 (s, 3H), 3.43 (s, 3H).

Alternatively,(2R,3R,4S,5S,6R)-2-(hydroxymethyl)-5,6-dimethoxytetrahydro-2H-pyran-3,4-diol(Compound A217) can be synthesized in the following manner:

Step 1: A solution of(2R,3R,4S,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yltrifluoromethanesulfonate (200 mg, 0.336 mmol) in MeOH (1 mL) is addedNaOMe (0.1 mL, 5.0 M in MeOH). The mixture was stirred at rt for 1 h.The resulting reaction mixture was monitored by TLC. When TLC showed thedisappearance of all starting material, the mixture was evaporated. Thecrude product was further purified by silica gel column chromatographyto give desired product (170 mg, 83% yield) as yellow oil. LC-MS (ESI)found: 501 [M+Na]⁺.

Step 2: To a solution of(2R,3S,4S,5S,6R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5,6-dimethoxytetrahydro-2H-pyran(35 mg, 0.063 mmol) in MeOH (3 mL) was added Pd/C (10 mg, 10% wt, 60%wet) at rt under H₂. The reaction was stirred at rt under a H₂ balloonfor 3 h. The resulting mixture was filtered and concentrated in vacuo togive2R,3R,4S,5S,6R)-2-(hydroxymethyl)-5,6-dimethoxytetrahydro-2H-pyran-3,4-diol(15 mg, 55% yield). ¹H NMR (400 MHz, Methanol-d₄) δ 4.33 (d, J=4.6 Hz,1H), 4.17 (p, J=5.0 Hz, 2H), 4.03-3.97 (m, 1H), 3.89 (t, J=4.9 Hz, 1H),3.79-3.68 (m, 2H), 3.44 (s, 3H), 3.43 (s, 3H). LC-MS (ESI) found: 231[M+Na]⁺

Synthesis 5-43. Preparation of(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-6-methoxy-5-morpholinotetrahydro-2H-pyran-3,4-diol(Compound A218)

Step 1:4-((2R,3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yl)morpholine(A218-1) was prepared following a similar procedure for(2R,3S,4R,5R,6R)-N-benzyl-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-amine(A214-4) in Synthesis 5-38. Yield: 550 mg, 62%. LC-MS (ESI) found: 534[M+H]⁺.

Step 2:(2R,3R,4R,5S,6R)-2-(hydroxymethyl)-6-methoxy-5-morpholinotetrahydro-2H-pyran-3,4-diol(A218) was prepared following a similar procedure for(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A214) in Synthesis 5-38. Yield: 6 mg, 30%. LC-MS (ESI) found: 264[M+H]⁺. ¹H NMR (400 MHz, CD₃OD): δ4.49 (d, J=1.9 Hz, 1H), 3.85-3.79 (m,2H), 3.78-3.74 (m, 1H), 3.74-3.66 (m, 4H), 3.60 (ddd, J=11.8, 6.0, 3.2Hz, 2H), 3.50 (s, 3H), 3.15 (t, J=20.8 Hz, 2H), 3.03-2.93 (m, 2H), 2.88(s, 1H).

Synthesis 5-44: Preparation ofN-((2R,3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)-N-methylacetamide(Compound A219)

Step 1: To a solution of(2R,3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxy-N-methyltetrahydro-2H-pyran-3-aminefrom Synthesis 5-41 (A216-1, 100 mg, 0.210 mmol) and TEA (64 mg, 0.630mmol) in DCM (5 mL) was added AcC1 (25 mg, 0.315 mmol) dropwise at 0° C.The reaction mixture was stirred at rt for 1.5 h. The resulting mixturewas diluted with DCM (10 mL), washed with H₂O (10 mL×2) and brine (10mL), dried over Na₂SO₄, filtered. The organic layer was concentrated invacuo to give a crude product, which was purified by flashchromatography (silica gel, 0˜80% EA in PE) to giveN-((2R,3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-2-methoxytetrahydro-2H-pyran-3-yl)-N-methylacetamide(A219-1). LC-MS (ESI) of both found: 520 [M+H]⁺.

Step 2:N-((2R,3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)-N-methylacetamide(A219) was prepared following a similar procedure for(2R,3R,4R,5S,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A214) in Synthesis 5-38. Yield: 7 mg, 32%. LC-MS (ESI) found: 250[M+H]⁺.

Synthesis 5-46. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(trifluoromethyl)tetrahydro-2H-pyran-3,4-diol(Compound A221)

Step 1: A 10 mL flame-dried round bottom flask was charged with5-(trifluoromethyl)-5H-dibenzo[b,d]thiophen-5-iumtrifluoromethanesulfonate (A126, 1.0 g, 2.49 mmol) and fac-Ir(ppy)₃ (8mg, 1.5 mol %). Then the flask was degassed and filled with argon forthree times.(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran(345 mg, 0.83 mmol) in DMA (10 mL) was added and the flask was sealed.The reaction mixture was stirred upon irradiation with blue bulbs (12 W*2; λ_(max)=465 nm) at room temperature. After 12 h, the reactionmixture was poured into water (15 mL), and then extracted with ethylacetate (10*3 mL).

The combined organic phase was washed with brine, dried over Na₂SO₄,filtered and concentrated to give a crude product, which was purified bycolumn chromatography (silica gel, 0-20% MeOH in DCM) to afford theproduct (A221-1, 100 mg, 25% yield). LC-MS (ESI) found: 507 [M+Na]⁺.

Step 2: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(trifluoromethyl)-3,4-dihydro-2H-pyran(A221-1, 50 mg, 0.10 mmol) in dry DCM (5 mL) at-78° C. under N₂atmosphere was added BCl₃ (1 mL, 1 M in DCM) slowly. After the addition,the reaction was stirred at room temperature for overnight. Onconsumption of starting material monitored by TLC, the reaction mixturewas quenched with 1 mL MeOH. The mixture was concentrated in vacuo togive(2R,3R,4R)-2-(hydroxymethyl)-5-(trifluoromethyl)-3,4-dihydro-2H-pyran-3,4-diol(A221-2). LC-MS (ESI) found: 213 [M-1]⁻.

Step 3: To a solution of(2R,3R,4R)-2-(hydroxymethyl)-5-(trifluoromethyl)-3,4-dihydro-2H-pyran-3,4-diol(A221-2, 20 mg, 0.10 mmol) in MeOH (5 mL) was added Pd/C (3 mg, 10% wt,60% wet). The reaction mixture was charged with H₂ for three times andstirred at rt for 16 h under H₂ atmosphere. The mixture was filtered andconcentrated in vacuo to give product (A221). LC-MS (ESI) found: 239[M+Na]⁺.

Alternatively,(2R,3R,4R)-2-(hydroxymethyl)-5-(trifluoromethyl)-3,4-dihydro-2H-pyran-3,4-diol(A221-2) can be synthesized in the following manner:

Step 1: A 10 mL flame-dried round bottom flask was charged with5-(trifluoromethyl)-5H-dibenzo[b,d]thiophen-5-iumtrifluoromethanesulfonate (996 mg, 2.48 mmol) and fac-Ir(ppy)₃ (8 mg,1.5 mol %). Then the flask was degassed and filled with argon for threetimes. (2R,3R,4R)-2-(acetoxymethyl)-3,4-dihydro-2H-pyran-3,4-diyldiacetate (225 mg, 0.82 mmol) in DMA (10 mL) was added and the flask wassealed. The reaction mixture was stirred upon irradiation with bluebulbs(12 W *2; λmax=465 nm) at room temperature. After 12 h, thereaction mixture was poured into water (15 mL), and then extracted withethyl acetate (10*3 mL). The combined organic phase was washed withbrine, dried (Na₂SO₄), concentrated and purified through columnchromatography (silica gel, 0-20% MeOH in DCM) to afford the crudeproduct (110 mg, purity: 80%). LC-MS (ESI) found: 341 [M+H]⁺.

Step 2: A solution of(2R,3R,4R)-2-(acetoxymethyl)-5-(trifluoromethyl)-3,4-dihydro-2H-pyran-3,4-diyldiacetate (30 mg, 0.08 mmol) in MeOH (0.6 mL, 13.9 mmol) was added NaOMe(4.76 mg, 0.088 mmol), the mixture was stirred at rt for 2 h. Themixture was acidification with AMBERLITE IR-120. The reaction wasfiltered and concentrated togive(2R,3R)-2-(hydroxymethyl)-5-(trifluoromethyl)-3,4-dihydro-2H-pyran-3,4-diol(A221-2, 18 mg, 95% yield). LC-MS (ESI) found: 237 [M+Na]⁺. ¹H NMR (400MHz, MeOD): δ 7.02 (s, 1H), 4.45 (d, J=3.7 Hz, 1H), 4.11-4.04 (m, 1H),3.98 (dd, J=4.4, 2.5 Hz, 1H), 3.89 (dd, J=11.9, 6.8 Hz, 1H), 3.79 (dd,J=11.9, 4.7 Hz, 1H).

Synthesis 5-47. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-morpholinotetrahydro-2H-pyran-3,4-diol(Compound A222)

Step 2: To a solution of(3S,4R,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-ol(100 mg, 0.23 mmol) in pyridine(2 mL) was added Tf₂O (324 mg, 1.15 mmol)at 0° C. After stirring at rt overnight, the mixture was concentratedand the residue was purified by flash chromatography (silica gel, 0-30%EtOAc in PE) to give(3S,4S,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yltrifluoromethanesulfonate (60 mg, 46% yield) as a colorless oil. LC-MS(ESI) found: 567 [M+H]⁺.

Step 3: A solution of(3S,4S,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yltrifluoromethanesulfonate (60 mg, 0.11 mmol) and morpholine (0.5 mL) inTHF (1 mL) was stirred at 80° C. overnight. The mixture was purified byprep-HPLC (Method A) to give4-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)morpholine(20 mg, 38% yield) as a colorless oil. LC-MS (ESI) found: 504 [M+H]⁺. ¹HNMR (400 MHz, CDCl₃): δ 7.38-7.26 (m, 15H), 4.84 (d, J=11.4 Hz, 1H),4.65 (d, J=11.4 Hz, 1H), 4.61 (t, J=6.0 Hz, 3H), 4.51 (d, J=12.1 Hz,1H), 4.20 (d, J=5.5 Hz, 1H), 4.11 (t, J=9.9 Hz, 2H), 3.93 (t, J=10.6 Hz,1H), 3.76-3.57 (m, 7H), 2.66 (d, J=47.9 Hz, 5H).

Step 4: A solution of4-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)morpholine(20 mg, 0.04 mmol) in MeOH (2 mL) were added Pd/C (5 mg, 10% wt, 60%wet) and HCl (0.1 mL, 1 M in H₂O). The mixture was stirred at rtovernight under a H₂ balloon. The mixture was filtered through a Celitepad, and the filtrate was concentrated to give(2R,3R,4R,5R)-2-(hydroxymethyl)-5-morpholinotetrahydro-2H-pyran-3,4-diolhydrochloride (A222, 6.5 mg, 70%) as a colorless oil. LC-MS (ESI) found:234[M+H]⁺. ¹H NMR (400 MHz, CD₃OD): δ4.44 (dd, J=14.3, 1.8 Hz, 1H), 4.16(dd, J=17.3, 6.6 Hz, 2H), 4.09 (dd, J=6.0, 2.8 Hz, 1H), 4.09-4.01 (m,2H), 3.80 (dd, J=10.0, 2.4 Hz, 2H), 3.77-3.72 (m, 2H), 3.71-3.62 (m,2H), 3.61-3.52 (m, 2H), 3.39 (dd, J=16.4, 7.1 Hz, 2H).

Synthesis 5-48. Preparation of3-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)oxazolidin-2-one(Compound A223)

Alternatively,3-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)oxazolidin-2-one(Compound A223) can be prepared in the same manner as Compound A180.

LC-MS (ESI) found: 234 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 4.41 (m, 1H),4.31 (m, 2H), 4.10 (dd, J=12.8, 2.3 Hz, 1H), 4.03 (m, 1H), 3.89-3.76 (m,3H), 3.75-3.63 (m, 3H), 3.45 (m, 1H).

Synthesis 5-49. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)imidazolidin-2-one(Compound A224)

Synthesis 5-50. Preparation of1-((1S,2R,3R,4S)-1-(aminomethyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)imidazolidine-2-thione(Compound A225)

Synthesis 5-51. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyrrolidine-2,5-dione(Compound A226)

Synthesis 5-52. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-1H-pyrrole-2,5-dione(Compound A227)

Synthesis 5-53. Preparation of3-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)thiazolidine-2,4-dione(Compound A228)

Synthesis 5-54. Preparation of3-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)oxazolidine-2,4-dione(Compound A229)

Synthesis 5-55. Preparation of2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)isoindoline-1,3-dione(Compound A230)

Synthesis 5-56. Preparation of2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)isoindolin-1-one(Compound A231)

Synthesis 5-57. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1H-imidazol-1-yl)tetrahydro-2H-pyran-3,4-diol(Compound A232)

Synthesis 5-58. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1H-pyrrol-1-yl)tetrahydro-2H-pyran-3,4-diol(Compound A234)

Synthesis 5-59. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyridin-2(1H)-one(Compound A235)

Synthesis 5-60. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyrimidin-2(1H)-one(Compound A236)

Synthesis 5-61. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyridin-4(1H)-one(Compound A237)

Synthesis 5-62. Preparation of(3S,4R,5R,6R)-6-(aminomethyl)-3-(piperidin-1-yl)tetrahydro-2H-pyran-2,4,5-triol(Compound A238)

Synthesis 5-63. Preparation of(3S,4R,5R,6R)-6-(aminomethyl)-3-morpholinotetrahydro-2H-pyran-2,4,5-triol(Compound A239)

Synthesis 5-64. Preparation of(3S,4R,5R,6R)-6-(aminomethyl)-3-thiomorpholinotetrahydro-2H-pyran-2,4,5-triol(Compound A240)

Synthesis 5-65. Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)azetidin-2-one(Compound A241)

Synthesis 5-66. Preparation of1-((3S,4R,5R,6R)-6-(aminomethyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-yl)tetrahydropyrimidin-2(1H)-one(Compound A242)

Synthesis 5-67. Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methanesulfinamide(Compound A243) andN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-1,1,1-trifluoromethanesulfinamide(Compound A244)

Synthesis 5-68. Alternative Preparation of(4aR,6R,7R,8R,8aS)-6-(aminomethyl)-7,8-dihydroxytetrahydro-1H,6H-pyrano[2,3-b][1,4]oxazin-2(3H)-one(Compound A245)

Synthesis 5-69. Preparation of(4aS,6R,7R,8R,8aS)-6-(aminomethyl)-7,8-dihydroxyhexahydro-1H,3H-pyrano[3,2-c][1,2,6]thiadiazine2,2-dioxide (Compound A246),(4aS,6R,7R,8R,8aS)-6-(aminomethyl)-7,8-dihydroxyhexahydro-1H-pyrano[3,2-d]pyrimidin-2(3H)-one(Compound A247), and(4aR,6R,7R,8R,8aS)-6-(aminomethyl)-7,8-dihydroxyhexahydropyrano[3,2-d][1,3]oxazin-2(1H)-one(Compound A248)

Synthesis 5-70. Preparation of((3aS,5R,6R,7R,7aS)-7-amino-5-(hydroxymethyl)-2-methyl-3a,6,7,7a-tetrahydro-5H-pyrano[3,2-d]oxazol-6-ol(Compound A249) and(3aS,5R,6R,7R,7aS)-7-amino-5-(hydroxymethyl)-2-(trifluoromethyl)-3a,6,7,7a-tetrahydro-5H-pyrano[3,2-d]oxazol-6-ol(Compound A250)

Synthesis 5-71. Preparation of(3aR,5R,6R,7R,7aS)-7-amino-5-(hydroxymethyl)-2-methyl-3,3a,5,6,7,7a-hexahydropyrano[2,3-d]imidazol-6-ol(Compound A251) and(3aR,5R,6R,7R,7aS)-7-amino-5-(hydroxymethyl)-2-(trifluoromethyl)-3,3a,5,6,7,7a-hexahydropyrano[2,3-d]imidazol-6-ol(Compound A252)

Synthesis 5-72. Preparation of1-((3aS,5R,6R,7R,7aS)-5-(aminomethyl)-6,7-dihydroxyhexahydropyrano[3,2-b]pyrrol-1(2H)-yl)-2,2,2-trifluoroethan-1-one(Compound A253)

Synthesis 5-73. Preparation of1-((4aR,6R,7R,8R,8aS)-6-(aminomethyl)-7,8-dihydroxyhexahydro-1H,6H-pyrano[2,3-b][1,4]oxazin-1-yl)ethan-1-one(Compound A254) and1-((4aR,6R,7R,8R,8aS)-6-(aminomethyl)-7,8-dihydroxyhexahydro-1H,6H-pyrano[2,3-b][1,4]oxazin-1-yl)-2,2,2-trifluoroethan-1-one(Compound A255)

Synthesis 5-74. Preparation of1-((3aS,4R,5aR,9aS,9bR)-4-(aminomethyl)-2,2,7-trimethylhexahydro-4H,9H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazin-9-yl)ethan-1-one(Compound A256) and1-((3aS,4R,5aR,9aS,9bR)-4-(aminomethyl)-2,2,7-trimethylhexahydro-4H,9H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazin-9-yl)-2,2,2-trifluoroethan-1-one(Compound A257)

Synthesis 5-75. Preparation of1-((3aS,4R,5aR,11aR,11bR)-4-(aminomethyl)-2,2-dimethyloctahydro-4H,11H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazocin-11-yl)ethan-1-one(Compound A258) and1-((3aS,4R,5aR,11aR,11bR)-4-(aminomethyl)-2,2-dimethyloctahydro-4H,11H-[1,3]dioxolo[4′,5′:4,5]pyrano[2,3-b][1,4]oxazocin-1l-yl)-2,2,2-trifluoroethan-1-one(Compound A259)

Synthesis 5-76. Preparation of1-((3aS,5R,6R,7R,7aR)-6,7-dihydroxy-5-(hydroxymethyl)hexahydropyrano[3,2-b]pyrrol-1(2H)-yl)ethan-1-one(Compound A260) and1-((2R,3R,4R,4aR,8aS)-3,4-dihydroxy-2-(hydroxymethyl)octahydro-5H-pyrano[3,2-b]pyridin-5-yl)ethan-1-one(Compound A261)

Synthesis 5-77. Preparation of1-((2R,3R,4R,4aR,9aS)-3,4-dihydroxy-2-(hydroxymethyl)octahydropyrano[3,2-b]azepin-5(2H)-yl)ethan-1-one(Compound A262)

Synthesis 5-78. Preparation of((3aR,4R,5aS,9aR,9bR)-2,2-dimethyl-8-oxooctahydro-4H-[1,3]dioxolo[4′,5′:4,5]pyrano[3,2-b]pyridin-4-yl)methylacetate (Compound A263)

Synthesis 5-79. Preparation ofN-((3aR,8S,8aR)-4-(hydroxymethyl)-2,2-dimethylhexahydro-4H-4,7-epoxy[1,3]dioxolo[4,5-d]azepin-8-yl)acetamide(Compound A264)

Synthesis 5-80. Preparation ofN-((3aR,4S,9R,9aR)-9-(hydroxymethyl)-2,2-dimethyloctahydro-5,9-epoxy[1,3]dioxolo[4,5-d]azocin-4-yl)acetamide(Compound A265)

Synthesis 5-81. Preparation ofN-((3aR,4S,9R,9aR)-9-(hydroxymethyl)-2,2-dimethylhexahydro-5H-5,9-epoxy[1,3]dioxolo[4,5-d]oxocin-4-yl)acetamide(Compound A266)

Synthesis 5-82. Compound A267, Compound A268, and Compound A269 can alsobe synthesized using Synthesis 5-79-5-81

Synthesis 5-82. General Synthesis to install R²

Synthesis 5-83. Alternative General Synthesis to install R²

Synthesis 5-84. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(isoxazol-5-ylamino)tetrahydro-2H-pyran-3,4-diol(Compound A272)

Synthesis 5-85. Preparation of(2R,3R,4R,5R)-5-((4,6-dichloro-1,3,5-triazin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A273)

Synthesis 5-86. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(thiazol-2-ylamino)tetrahydro-2H-pyran-3,4-diol(Compound A274)

Synthesis 5-87. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-((3-(trifluoromethyl)pyridin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol.(Compound A275)

Synthesis 5-88. Preparation of(3aR,4R,8R,8aR)-8-azido-4-(azidomethyl)-2,2-dimethylhexahydro-4H-4,7-epoxycyclohepta[d][1,3]dioxole(Compound A276)

Synthesis 5-89. Preparation of Compound A277 and Compound A278

Alternatively, Compound A279 can be synthesized if

is used instead of

in the Schotten Bauman reaction step.

Synthesis 5-90. Preparation of1-((3S,4R,5R,6R)-6-(aminomethyl)-2,4,5-trihydroxytetrahydro-2H-pyran-3-yl)guanidine(Compound A280) And (Compound A281)

Synthesis 5-91. Preparation of Compound A282

Example 6. Synthesis of Degraders Synthesis 6-1. Preparation ofBidentate Fumaramide OPT-3 (Compound 10)

The —NH(OPT-3)C(O)CH₂SH is generated in situ from NH₂OH treatment ofSATA-(N-succinimidyl S-acetylthioacetate)-OPT-3.Synthesis 6-2. General Synthesis of Bidentate Fumaramide OPT-3conjugate-sulfoximine Compounds

The —NH(OPT-3)C(O)CH₂SH is generated in situ from NH₂OH treatment ofSATA-(N-succinimidyl S-acetylthioacetate)-OPT-3. Compound 12 can besynthesized using the procedure of Synthesis 2-58 with MeLi.

Synthesis 6-3. Preparation of Compound 13

Compound 13-1 is synthesized from ASGPR Ligand A245 from Synthesis5-68^(PGP)-7⁸⁶,C₃

Synthesis 6-4. Preparation of Compound 14

Compound 14-1 is synthesized from ASGPR Ligand A253 from Synthesis 5-72

Synthesis 6-5. Preparation of Compound 15 and Compound 16

Synthesis 6-6. Preparation of Compound 17

Synthesis 6-7. Preparation of Compound 18

Synthesis 6-8. Preparation of Compound 19

Example 7. Synthesis and Installation of Linkers

Linker^(B) can be synthesized from any chemical moiety containing atleast two reactive sites for bond formation. For example, a compound ofthe present invention containing Linker^(B) can be synthesized from:

Linker^(C) can be synthesized from any chemical moiety containing atleast three reactive sites for bond formation. For example, a compoundof the present invention containing Linker^(C) can be synthesized from:

Linker^(D) can be synthesized from any chemical moiety containing atleast three reactive sites for bond formation. For example, a compoundof the present invention containing Linker^(D) can be synthesized from:

Synthesis 7-1. Attachment of triazole-containing alkyl and polyethyleneglycol Linkers

Synthesis 7-2. Attachment of1-(3,4-dihydroxybenzyl)-1H-pyrrole-2,5-dione Linker

Synthesis 7-3. Attachment of1-(carboxymethyl)-5-hydroxy-1H-indole-3-carboxylic acid Linker

Synthesis 7-4. Attachment of pyrrolidine-3,4-diamine Linker

Synthesis 7-5. Attachment of cyclobutane-1,3-diol-containing alkyl andpolyethylene glycol LinkersFor linear alkyl:

Linker^(C):

For polyethylene glycol:

Linker^(C):

Linker^(D):

Alternatively, for linear alkyl:

Linker^(C):

Linker^(D):

Alternatively, for polyethylene glycol:

Linker^(C):

Linker^(D):

Example 8. Additional Synthetic Procedures

Preparation of(1S,2R,3R,4R,5S)-4-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(Compound A287)

Step 1: A solution of(1S,2R,3R,4R,5S)-4-amino-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(50 mg, 0.26 mmol), 3,5-dichloro-1,2,4-thiadiazole (121 mg, 0.78 mmol)and DIPEA (169 mg, 1.3 mmol) in i-PrOH (1 mL) was stirred at 80° C.overnight.

The mixture was concentrated and the residue was purified by prep togive(1S,2R,3R,4R,5S)-4-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(A287, 60 mg, 74% yield) as a yellow solid. LC-MS (ESI) found: 310[M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 5.38 (d, J=0.9 Hz, 1H), 3.97-3.86 (m,3H), 3.83 (d, J=11.4 Hz, 1H), 3.80-3.75 (m, 2H), 3.71 (d, J=8.0 Hz, 1H).

The following compounds below were prepared according to the sameprocedure as A287:

Starting ID Characterization data Material

Yield: 3.5 mg, 3.2%, white solid. LC-MS (ESI) found: 354 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 5.38 (s, 1H), 3.91 (dd, J = 12.5, 7.7 Hz, 3H),3.83 (d, J = 11.4 Hz, 1H), 3.80- 3.75 (m, 2H), 3.71 (d, J = 8.0 Hz, 1H).

Yield: 21 mg, 48%, white solid. LC-MS (ESI) found: 338 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 8.52 (d, J = 4.9 Hz, 1H), 6.90 (dd, J = 14.0, 5.8 Hz,1H), 5.39 (t, J = 14.0 Hz, 1H), 4.20 (d, J = 9.7 Hz, 1H), 3.96-3.78 (m,5H), 3.71 (d, J = 7.9 Hz, 1H).

Yield: 6 mg, 20%, white solid. LC-MS (ESI) found: 276 [M + H]⁺, ¹H NMR(400 MHz, MeOD): δ 7.85 (s, 1H), 5.40 (d, J = 1.0 Hz, 1H), 3.92 (dd, J =11.8, 7.8 Hz, 3H), 3.83-3.77 (m, 3H), 3.71 (d, J = 8.0 Hz, 1H).

Yield: 20 mg, 22%, white solid. LC-MS (ESI) found: 343 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.48 (dd, J = 2.5, 1.5 Hz, 1H), 7.01 (dd, J = 4.5,1.5 Hz, 1H), 6.62 (dd, J = 4.5, 2.6 Hz, 1H), 5.42 (d, J = 1.4 Hz, 1H),4.50 (dd, J = 9.9, 1.1 Hz, 1H), 4.03 (dd, J = 9.9, 4.3 Hz, 1H), 3.96 (d,J = 7.5 Hz, 1H), 3.94 (s, 1H), 3.84 (dd, J = 9.6, 6.1 Hz, 2H), 3.74 (d,J = 7.9 Hz, 1H).

Yield: 16 mg, 25%, white solid. LC-MS (ESI) found: 334 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.06 (s, 1H), 5.39 (s, 1H), 4.15 (d, J = 9.2 Hz, 1H),3.96-3.91 (m, 2H), 3.85- 3.76 (m, 3H), 3.70 (d, J = 7.9 Hz, 1H), 3.35(s, 3H).

Yield: 2 mg, 6%, white solid. LC-MS (ESI) found: 334 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.23 (s, 1H), 5.35 (s, 1H), 4.30 (d, J = 9.9 Hz, 1H),3.98-3.88 (m, 5H), 3.85- 3.75 (m, 3H), 3.70 (d, J = 7.9 Hz, 1H).

Preparation of2-(((1S,2R,3R,4R,5S)-2,3-dihydroxy-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octan-4-yl)amino)-6-methoxypyrimidine-4-carbonitrile(Compound A294)

Step 1: To a solution of(1S,2R,3R,4R,5S)-4-((4-chloro-6-methoxypyrimidin-2-yl)amino)-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octane-2,3-diol(A292, 30 mg, 0.09 mmol) in DMF (2 mL) was added dppf (14 mg, 0.024mmol), Pd₂(dba)₃ (6 mg, 0.006 mmol) and Zn(CN)₂ (14 mg, 0.12 mmol) atrt. After the addition was complete, the mixture was stirred at 120° C.under N₂ atmosphere overnight. On consumption of starting material (LCMSmonitoring), the mixture was filtered, concentrated, and purified byflash chromatography (silica gel, 0-10% MeOH in DCM) to give2-(((1S,2R,3R,4R,5S)-2,3-dihydroxy-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octan-4-yl)amino)-6-methoxypyrimidine-4-carbonitrile(2.7 mg, 9% yield) as white solid. LC-MS (ESI) found: 325 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.46 (s, 1H), 5.35 (s, 1H), 4.17 (d, J=8.6 Hz, 1H),3.94 (d, J=11.3 Hz, 5H), 3.87-3.76 (m, 3H), 3.70 (d, J=7.9 Hz, 1H).

Reduced Galactose Series:

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A295)

To a mixture of(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diolhydrochloride (50 mg, 0.25 mmol) in NMIP (2.0 mL) was added2-chloro-4-(trifluoromethyl)pyrimidine (168 mg, 0.92 mmol) and TEA (124mg, 1.2 mmol) at rt under N₂. After stirring at 120° C. overnight, themixture was concentrated and purified by prep-TLC to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(3 mg, 3.2% yield) as white solid. LC-MS (ESI) found: 310 [M+H]⁺. ¹H NMR(400 MHz, CD₃OD): δ8.51 (d, J=4.8 Hz, 1H), 6.89 (d, J=4.9 Hz, 1H), 4.36(td, J=10.6, 5.3 Hz, 1H), 4.11 (dd, J=10.9, 5.2 Hz, 1H), 3.91 (d, J=2.6Hz, 1H), 3.78-3.65 (m, 3H), 3.45 (ddd, J=6.9, 5.0, 1.0 Hz, 1H), 3.16 (t,J=10.9 Hz, 1H).

The following compounds below were made using the method described forA295:

Starting ID Characterization data material

Yield: 2.3 mg, 5%, white solid. LC-MS (ESI) found: 309 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.56 (d, J = 1.4 Hz, 1H), 6.48 (d, J = 1.4 Hz, 1H),4.23 (td, J = 10.5, 5.2 Hz, 1H), 4.08 (dd, J = 11.0, 5.2 Hz, 1H), 3.89(d, J = 2.8 Hz, 1H), 3.71 (dt, J = 11.4, 5.3 Hz, 2H), 3.55 (dd, J =10.5, 3.2 Hz, 1H), 3.46-3.42 (m, 1H), 3.08 (t, J = 10.8 Hz, 1H).

Yield: 1.4 mg, 3%, white solid LC-MS (ESI) found: 309 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.62 (s, 2H), 3.95 (dd, J = 11.3, 5.1 Hz, 1H), 3.89(d, J = 2.9 Hz, 1H), 3.85-3.77 (m, 1H), 3.77-3.66 (m, 2H), 3.51 (dd, J =10.0, 3.2 Hz, 1H), 3.44-3.40 (m, 1H), 3.10 (t, J = 11.0 Hz, 1H).

Yield: 2.7 mg, 5%, white solid. LC-MS (ESI) found: 309 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.93 (s, 1H), 6.90 (s, 1H), 3.99- 3.91 (m, 3H),3.76-3.68 (m, 3H), 3.48-3.45 (m, 1H), 3.28-3.23 (m, 1H).

Yield: 65 mg, 66%, oil. LC-MS (ESI) found: 326 [M + H]+. ¹H NMR (400MHz, D₂O): δ 4.10-4.02 (m, 1H), 3.92 (d, J = 1.4 Hz, 1H), 3.80-3.59 (m,4H), 3.57-3.50 (m, 1H), 3.23 (t, J = 11.0 Hz, 1H).

Yield: 15.3 mg, 32%, white solid. LC-MS (ESI) found: 316 [M + H]⁺ . ¹HNMR (400 MHz, MeOD): δ 4.15 (dd, J = 11.0, 5.1 Hz, 1H), 4.02 (s, 1H),3.90 (d, J = 2.5 Hz, 1H), 3.71 (ddd, J = 16.4, 11.4, 6.0 Hz, 2H), 3.61(dd, J = 10.3, 3.2 Hz, 1H), 3.47-3.41 (m, 1H), 3.20 (t, J = 10.9 Hz,1H).

Yield: 6.7 mg, 7%, orange solid. LC-MS (ESI) found: 310 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 8.00 (s, 1H), 6.55 (s, 1H), 4.38 (s, 1H), 4.02 (s,1H), 3.82 (d, J = 2.7 Hz, 1H), 3.63 (ddd, J = 16.4, 11.4, 6.0 Hz, 2H),3.54 (dd, J = 10.5, 3.0 Hz, 1H), 3.38-3.33 (m, 1H), 3.03 (t, J = 9.8 Hz,1H).

Yield: 2.9 mg, 3.4%, solid. LC/MS ESI(m/z): 309 [M + H]⁺. ¹H NMR (400MHz, MeOD) δ 7.52 (t, J = 7.9 Hz, 1H), 6.88 (d, J = 7.2 Hz, 1H), 6.72(d, J = 8.5 Hz, 1H), 4.29 (td, J = 10.5, 5.2 Hz, 1H), 4.18 (dd, J =10.9, 5.2 Hz, 1H), 3.90 (d, J = 2.9 Hz, 1H), 3.72 (ddd, J = 16.3, 11.4,6.0 Hz, 2H), 3.60 (dd, J = 10.4, 3.2 Hz, 1H), 3.49-3.42 (m, 1H), 3.11(t, J = 10.7 Hz, 1H).

Yield: 8.9 mg, 30%, white solid. LC/MS ESI(m/z): 306 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.04 (s, 1H), 4.31 (td, J = 10.6, 5.2 Hz, 1H), 4.11(dd, J = 10.9, 5.1 Hz, 1H), 3.90 (d, J = 2.9 Hz, 4H), 3.73 (d, J = 7.0Hz, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.61 (dd, J = 10.5, 2.7 Hz,1H), 3.46-3.41 (m, 1H), 3.13 (t, J = 10.8 Hz, 1H).

Yield: 5 mg, 5%, solid. LC/MS ESI(m/z): 315 [M + H]⁺. ¹H NMR (400 MHz,MeOD): δ 7.47 (dd, J = 2.6, 1.5 Hz, 1H), 6.90 (dd, J = 4.4, 1.4 Hz, 1H),6.61 (dd, J = 4.4, 2.6 Hz, 1H), 4.69 (td, J = 10.8, 5.3 Hz, 1H), 4.11(dd, J = 11.0, 5.3 Hz, 1H), 3.93 (d, J = 2.8 Hz, 1H), 3.79- 3.68 (m,3H), 3.47 (ddd, J = 6.8, 5.0, 0.9 Hz, 1H), 3.24 (t, J = 10.9 Hz, 1H).

Yield: 6.0 mg, 7%, a white solid. LC-MS (ESI) found: 316 [M + H]⁺. ¹HNMR (400 MHz, MeOD) 8 4.20 (dd, J = 11.0, 5.2 Hz, 1H), 4.07 (td, J =10.5, 5.2 Hz, 1H), 3.90 (d, J = 2.9 Hz, 1H), 3.75 (dd, J = 11.4, 7.1 Hz,1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.61 (dd, J = 10.3, 3.1 Hz, 1H),3.47-3.43 (m, 1H), 3.20 (t, J = 10.9 Hz, 1H).

Yield: 62.8 mg, 68%, oil. LC-MS (ESI) found: 315 [M + H]⁺. ¹H NMR (400MHz, MeOD): δ 7.08 (s, 1H), 4.16 (dd, J = 11.0, 5.2 Hz, 1H), 4.04 (m,1H), 3.89 (d, J = 2.9 Hz, 1H), 3.74 (dd, J = 11.4, 7.0 Hz, 1H), 3.67(dd, J = 11.3, 4.9 Hz, 1H), 3.57 (dd, J = 10.8, 3.6 Hz, 1H), 3.46-3.40(m, 1H), 3.14 (t, J = 10.8 Hz, 1H).

Yield: 3.8 mg, 11%, white solid. LC-MS (ESI) found: 343 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.98 (d, J = 6.2 Hz, 1H), 6.98 (d, J = 6.2 Hz, 1H),4.07-3.98 (m, 2H), 3.91 (dd, J = 3.1, 0.8 Hz, 1H), 3.78-3.65 (m, 3H),3.50-3.44 (m, 1H), 3.23 (s, 1H).

Yield: 5.3 mg, 6.9%, white solid. LC-MS (ESI) found: 310 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.10 (s, 1H), 8.02 (s, 1H), 4.34 (td, J = 10.6,5.1 Hz, 1H), 4.09 (dd, J = 11.0, 5.2 Hz, 1H), 3.91 (d, J = 2.9 Hz, 1H),3.76 (dd, J = 11.4, 7.1 Hz, 1H), 3.69 (dd, J = 11.4, 5.0 Hz, 1H), 3.61(dd, J = 10.5, 3.2 Hz, 1H), 3.50-3.40 (m, 1H), 3.11 (t, J = 10.9 Hz,1H).

Yield: 8.1 mg, 31%, yellow solid. LC-MS (ESI) found: 267 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.46 (t, J = 7.1 Hz, 1H), 6.94 (d, J = 4.7 Hz,1H), 4.44-4.18 (m, 1H), 4.06 (dd, J = 11.0, 5.2 Hz, 1H), 3.91 (d, J =3.0 Hz, 1H), 3.79-3.73 (m, 1H), 3.72-3.68 (m, 1H), 3.65 (dd, J = 10.5,3.2 Hz, 1H), 3.48-3.43 (m, 1H), 3.16 (t, J = 10.9 Hz, 1H).

Yield: 50 mg, 65%, white solid. LC-MS(ESI) found 310 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 8.50 (s, 1H), 6.84 (s, 1H), 4.53 (s, 1H), 4.10 (s,1H), 3.91 (d, J = 2.7 Hz, 1H), 3.76 (dd, J = 11.4, 7.0 Hz, 1H), 3.69(dd, J = 11.4, 5.0 Hz, 1H), 3.62 (dd, J = 10.5, 3.2 Hz, 1H), 3.47-3.43(m, 1H), 3.14 (d, J = 10.7 Hz, 1H).

Yield: 2.1 mg, 1%, white solid. LC-MS (ESI) found: 318 [M + H]⁺. ¹H NMR(400 MHz, CD3OD): δ 8.29 (d, J = 5.3 Hz, 1H), 8.20- 8.05 (m, 2H), 7.47(dd, J = 5.1, 1.9 Hz, 3H), 7.13 (d, J = 5.3 Hz, 1H), 4.44 (s, 1H), 4.23(dd, J = 11.0, 5.2 Hz, 1H), 3.93 (d, J = 2.7 Hz, 1H), 3.82-3.66 (m, 3H),3.52-3.45 (m, 1H), 3.20 (t, J = 10.8 Hz, 1H).

Yield: 3.3 mg, 5.1%, white solid. LC-MS (ESI) found: 267 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.10 (s, 1H), 8.02 (s, 1H), 4.34 (td, J = 10.6,5.1 Hz, 1H), 4.09 (dd, J = 11.0, 5.2 Hz, 1H), 3.91 (d, J = 2.9 Hz, 1H),3.76 (dd, J = 11.4, 7.1 Hz, 1H), 3.69 (dd, J = 11.4, 5.0 Hz, 1H), 3.61(dd, J = 10.5, 3.2 Hz, 1H), 3.50-3.40 (m, 1H), 3.11 (t, J = 10.9 Hz,1H).

Yield: 8.7 mg, 11%, white solid. LC-MS (ESI) found: 310 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.58 (d, J = 9.4 Hz, 1H), 7.02 (d, J = 9.4 Hz, 1H),4.44 (d, J = 4.4 Hz, 1H), 4.18 (dd, J = 11.0, 5.2 Hz, 1H), 3.92 (d, J =2.8 Hz, 1H), 3.77 (dd, J = 11.4, 7.1 Hz, 1H), 3.70 (dd, J = 11.4, 5.0Hz, 1H), 3.66 (dd, J = 10.4, 3.2 Hz, 1H), 3.51-3.44 (m, 1H), 3.18 (t, J= 10.9 Hz, 1H).

Yield: 9.1 mg, 12.8%, white solid. LC-MS (ESI) found: 310 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.27 (s, 1H), 7.94 (d, J = 1.0 Hz, 1H), 4.40 (td,J = 10.6, 5.1 Hz, 1H), 4.10 (dd, J = 11.0, 5.2 Hz, 1H), 3.91 (d, J = 2.8Hz, 1H), 3.72 (ddd, J = 16.3, 11.4, 6.0 Hz, 2H), 3.63 (dd, J = 10.5, 3.2Hz, 1H), 3.51-3.40 (m, 1H), 3.14 (t, J = 10.9 Hz, 1H).

Yield: 10.1 mg, 13%, white solid. LC-MS (ESI) found: 310 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.11 (s, 1H), 7.99 (s, 1H), 4.34 (td, J = 10.5,5.2 Hz, 1H), 4.14 (dd, J = 11.0, 5.2 Hz, 1H), 3.92 (d, J = 3.1 Hz, 1H),3.76 (dd, J = 11.4, 7.1 Hz, 1H), 3.71-3.62 (m, 2H), 3.49-3.44 (m, 1H),3.12 (t, J = 10.8 Hz, 1H).

Yield: 10 mg, 38%, yellow solid. LC-MS (ESI) found: 276[M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.57 (d, J = 9.3 Hz, 1H), 7.15 (d, J = 9.3 Hz, 1H),4.41 (ddd, J =19.4, 11.3, 5.9 Hz, 2H), 3.97-3.72 (m, 3H), 3.39 (dd, J =10.1, 3.3 Hz, 1H), 3.15-3.05 (m, 1H), 2.97 (td, J = 10.6, 5.1 Hz, 1H)

Yield: 4.1 mg, 6%, white solid. LC-MS (ESI) found: 276 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.85 (d, J = 2.3 Hz, 1H), 6.43 (d, J = 3.5 Hz, 1H),4.44 (dd, J = 16.4, 9.5 Hz, 1H), 4.07 (dd, J = 6.9, 3.8 Hz, 1H), 3.90(d, J = 2.8 Hz, 1H), 3.75 (dd, J = 11.4, 7.1 Hz, 1H), 3.68 (dd, J =11.4, 5.0 Hz, 1H), 3.58 (dd, J = 10.5, 3.2 Hz, 1H), 3.46-3.39 (m, 1H),3.10 (t, J = 10.6 Hz, 1H).

Yield: 9.8 mg, 16%, white solid. LC-MS (ESI) found: 242 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 8.26 (d, J = 4.7 Hz, 2H), 6.61 (t, J = 4.8 Hz, 1H),4.30 (td, J = 10.6, 5.2 Hz, 1H), 4.11 (dd, J = 11.0, 5.2 Hz, 1H), 3.90(d, J = 2.7 Hz, 1H), 3.76 (dd, J = 11.4, 7.0 Hz, 1H), 3.69 (dd, J =11.4, 5.0 Hz, 1H), 3.63 (dd, J = 10.5, 3.2 Hz, 1H), 3.45 (dd, J = 6.5,5.4 Hz, 1H), 3.15 (t, J =10.8 Hz, 1H).

Yield: 0.7 mg, 2.4%, white solid. LC-MS (ESI) found: 325 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.62 (d, J = 6.1 Hz, 2H), 8.11 (d, J = 6.2 Hz,2H), 4.23 (dd, J = 11.0, 5.2 Hz, 1H), 4.16-4.06 (m, 1H), 3.92 (d, J =3.0 Hz, 1H), 3.77 (dd, J = 11.4, 7.0 Hz, 1H), 3.70 (dd, J = 11.4, 5.1Hz, 1H), 3.65 (dd, J = 10.3, 3.1 Hz, 1H), 3.51-3.44 (m, 1H), 3.23 (t, J= 10.9 Hz, 1H).

Yield: 3.1 mg, 4.1%, white solid. LC-MS (ESI) found: 301 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.67 (s, 1H), 4.45 (dd, J = 10.5, 5.1 Hz, 1H),4.05 (dd, J = 11.0, 5.1 Hz, 1H), 3.90 (d, J = 2.9 Hz, 2H), 3.75 (dd, J =11.4, 7.1 Hz, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.58 (dd, J = 10.5,3.2 Hz, 1H), 3.48- 3.40 (m, 1H), 3.11 (t, J = 10.9 Hz, 1H).

Yield: 19.1 mg, 28%, white solid. LC-MS (ESI) found: 276 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.22 (s, 1H), 6.55 (s, 1H), 4.44 (dd, J = 12.1,4.9 Hz, 1H), 4.05 (dd, J = 6.1,2.4 Hz, 1H), 3.90 (d, J = 3.1 Hz, 1H),3.75 (dd, J = 11.4, 7.0 Hz, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.58(dd, J = 10.4, 3.0 Hz, 1H), 3.44 (dd, J = 6.5, 5.5 Hz, 1H), 3.12 (t, J =10.0 Hz, 1H).

Yield: 3.7 mg, 9%, white solid. LC-MS (ESI) found: 365[M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.92 (s, 1H), 7.51 (s, 2H), 4.23 (ddd, J = 15.7,11.7, 5.3 Hz, 2H), 3.92 (d, J = 2.5 Hz, 1H), 3.73 (ddd, J =16.3, 11.4,6.0 Hz, 2H), 3.62 (dd, J = 10.1, 3.2 Hz, 1H), 3.46 (ddd, J = 7.1, 5.3,3.8 Hz, 1H), 3.24-3.15 (m, 1H).

Yield: 8.3 mg, 20%, white solid. LC-MS (ESI) found: 272 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.93 (d, J = 5.9 Hz, 1H), 6.06 (d, J = 5.9 Hz, 1H),4.29 (td, J = 10.5, 5.1 Hz, 1H), 4.14 (dd, J = 10.9, 5.2 Hz, 1H), 3.91(s, 3H), 3.90 (s, 1H), 3.76 (dd, J = 11.4, 7.0 Hz, 1H), 3.69 (dd, J =11.4, 5.0 Hz, 1H), 3.63 (dd, J = 10.5, 3.2 Hz, 1H), 3.47-3.41 (m, 1H),3.15 (t, J = 10.8 Hz, 1H).

Yield: 5.4 mg, 10%, white solid. LC-MS (ESI) found: 344 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 8.46 (s, 1H), 4.32 (s, 1H), 4.07 (dd, J = 10.9, 5.1Hz, 1H), 3.90 (d, J = 2.8 Hz, 1H), 3.75 (dd, J = 11.4, 7.1 Hz, 1H),3.71-3.67 (m, 1H), 3.65 (d, J = 8.5 Hz, 1H), 3.47-3.41 (m, 1H), 3.16 (t,J = 10.9 Hz, 1H).

Yield: 8.3 mg, 11%, white solid. LC-MS (ESI) found: 303 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 4.39 (td, J = 10.7, 5.3 Hz, 1H), 4.08 (dd, J = 11.0,5.3 Hz, 1H), 4.08 (dd, J = 11.0, 5.3 Hz, 1H),, 4.01 (s, 1H), 3.95 (s,3H, 3.90 (s, 3H), 3.75 (dd, J = 11.4, 7.0 Hz, 1H), 3.71-3.66 (m, 1H),3.63 (dd, J = 10.6, 3.2 Hz, 1H), 3.47-3.40 (m, 1H), 3.16 (t, J = 10.9Hz, 1H).

Yield: 3.0 mg, 10%, white solid. LC-MS (ESI) found: 297 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.55 (s, 1H), 4.53-4.40 (m, 1H), 4.12 (dd, J = 10.9,5.3 Hz, 1H), 3.91 (dd, J = 8.4, 4.7 Hz, 6H), 3.75 (dd, J = 11.4, 7.1 Hz,1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.60 (dd, J = 9.9, 2.4 Hz, 1H),3.44 (dd, J = 8.7, 3.3 Hz, 2H), 3.11 (t, J = 10.7 Hz, 1H).

Yield: 3.0 mg, 1%, white solid. ¹H NMR (400 MHz, MeOD): δ 6.55 (s, 1H),4.53-4.40 (m, 1H), 4.12 (dd, J = 10.9, 5.3 Hz, 1H), 3.91 (dd, J = 8.4,4.7 Hz, 6H), 3.75 (dd, J = 11.4, 7.1 Hz, 1H), 3.68 (dd, J = 11.4, 5.0Hz, 1H), 3.60 (dd, J = 9.9, 2.4 Hz, 1H), 3.44 (dd, J = 8.7, 3.3 Hz, 2H),3.11 (t, J = 10.7 Hz, 1H).

Yield: 5.0 mg, 21%, white solid. LC-MS (ESI) found: 344 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.76 (s, 1H), 4.49 (td, J = 10.7, 5.2 Hz, 1H), 4.09(dd, J = 11.0, 5.2 Hz, 1H), 3.91 (d, J = 2.8 Hz, 1H), 3.76 (dd, J =11.4, 7.1 Hz, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.61 (dd, J = 10.6,3.1 Hz, 1H), 3.49- 3.42 (m, 1H), 3.12 (t, J = 10.9 Hz, 1H).

Yield: 3.1 mg, 4%, white solid. LC-MS (ESI) found: 306 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.04 (s, 1H), 4.31 (td, J = 10.5, 5.3 Hz, 1H), 4.11(dd, J = 10.9, 5.1 Hz, 1H), 3.90 (d, J = 2.8 Hz, 4H), 3.75 (dd, J =11.4, 7.0 Hz, 1H), 3.68 (dd, J = 11.3, 5.0 Hz, 1H), 3.61 (dd, J = 10.2,3.2 Hz, 1H), 3.46-3.40 (m, 1H), 3.13 (t, J = 10.8 Hz, 1H).

Yield: 8.0 mg, 6%, white solid. LC-MS (ESI) found: 307 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 4.48-4.37 (m, 1H), 4.10-4.03 (m, 1H), 4.02-3.95 (m,3H), 3.92 (t, J = 3.3 Hz, 1H), 3.79-3.61 (m, 3H), 3.48-3.42 (m, 1H),3.19 (td, J =11.0, 5.8 Hz, 1H).

Yield: 4.5 mg, 5%, white solid. LC-MS (ESI) found: 322 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.20 (s, 1H), 4.44 (dd, J = 27.9, 14.4 Hz, 1H), 4.10(dd, J = 15.2, 7.8 Hz, 1H), 3.90 (d, J = 2.6 Hz, 1H), 3.75 (dd, J =11.3, 7.1 Hz, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.58 (dd, J = 10.5,3.2 Hz, 1H), 3.43 (dd, J = 6.6, 5.5 Hz, 1H), 3.12 (t, 1H), 2.50 (s, 3H).

Yield: 2.0 mg, 3%, white solid. LC-MS (ESI) found: 266 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.47 (dd, J = 8.7, 7.2 Hz, 1H), 6.96 (dd, J = 7.1,0.6 Hz, 1H), 6.76 (d, J = 8.7 Hz, 1H), 4.32 (td, J = 10.6, 5.2 Hz, 1H),4.12 (dd, J = 11.0, 5.2 Hz, 1H), 3.90 (d, J = 2.7 Hz, 1H), 3.76 (dd, J =11.4, 7.1 Hz, 1H), 3.68 (dd, J = 11.4, 5.0 Hz, 1H), 3.58 (dd, J = 10.5,3.2 Hz, 1H), 3.44 (dd, J = 6.6, 5.5 Hz, 1H), 3.08 (t, J = 10.8 Hz, 1H).

Yield: 6.0 mg, 8%, white solid. LC-MS (ESI) found: 297 [M + H]⁺. ¹H NMR(400 MHz, CD₃0D): δ 5.97 (s, 1H), 4.36 (dd, J = 30.8, 19.2 Hz, 1H), 4.03(dd, J = 10.9, 4.6 Hz, 1H), 3.89 (d, J = 3.3 Hz, 4H), 3.75 (dd, J =11.4, 7.1 Hz, 1H), 3.67 (dd, J = 11.4, 5.0 Hz, 1H), 3.56 (dd, J = 10.4,3.1 Hz, 1H), 3.43 (dd, J = 6.6, 5.5 Hz, 1H), 3.10 (t, J = 10.8 Hz, 1H).

Yield: 8.0 mg, 8%, white solid. LC-MS (ESI) found: 324 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 6.82 (s, 1H), 4.36 (s, 1H), 4.12 (dd, J = 11.0, 5.2Hz, 1H), 3.90 (d, J = 2.9 Hz, 1H), 3.75 (dd, J = 11.4, 7.1 Hz, 1H),3.71- 3.61 (m, 2H), 3.47-3.42 (m, 1H), 3.15 (t, J = 10.8 Hz, 1H), 2.40(s, 3H).

Yield: 26 mg, 36%, white solid. LC-MS (ESI) found: 292 [M + H]⁺. ¹H NMR(400 MHz, CD₃OD): δ 8.42 (d, J = 4.9 Hz, 1H), 6.81 (d, J = 4.9 Hz, 1H),6.42 (t, J = 55.0 Hz, 1H), 4.36 (td, J = 10.6, 5.2 Hz, 1H), 4.11 (dd, J= 11.0, 5.2 Hz, 1H), 3.91 (d, J = 2.9 Hz, 1H), 3.75 (dd, J = 11.4, 7.0Hz, 1H), 3.71-3.60 (m, 2H), 3.50-3.40 (m, 1H), 3.16 (t, J = 10.9 Hz,1H).

Yield: 2.3 mg, 9%, white solid. LC-MS (ESI) found: 288 [M + H]⁺. ¹H NMR(400 MHz, CD₃OD): δ 7.87 (d, J = 3.7 Hz, 1H), 6.51 (d, J = 5.4 Hz, 1H),4.46-4.26 (m, 1H), 4.14 (dd, J = 11.0, 5.2 Hz, 1H), 3.90 (d, J = 2.8 Hz,1H), 3.76 (dd, J = 11.4, 7.1 Hz, 1H), 3.69 (dd, J = 11.4, 5.0 Hz, 1H),3.63 (dd, J = 10.5, 3.2 Hz, 1H), 3.50-3.42 (m, 1H), 3.15 (t, J = 10.8Hz, 1H), 2.51 (s, 3H).

Preparation of6-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-1,3,5-triazine-2,4(1H,3H)-dione(Compound A337)

It was prepared according to the procedure same as that for A112. Yield:1.5 mg, 2%, white solid. ¹H NMR (400 MHz, D₂O): δ4.73 (d, J=3.1 Hz, 2H),4.42 (dt, J=9.6, 3.2 Hz, 1H), 4.21 (dd, J=12.7, 3.0 Hz, 1H), 4.06 (td,J=6.7, 2.5 Hz, 1H), 3.76-3.66 (m, 3H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-(4-methylpiperazin-1-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A338)

Step 1: To a mixture of(2R,3R,4R,5S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A104, 100 mg, 0.36 mmol) in i-PrOH (3.0 mL) was added1-methylpiperazine (107 mg, 1.07 mmol) and DIEA (0.25 mL, 1.44 mmol) atrt under N₂. After stirring at 120° C. overnight, the mixture wasconcentrated and purified by prep-TLC to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-(4-methylpiperazin-1-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(1.8 mg, 6% yield) as a white solid. LC-MS (ESI) found: 346 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.47 (s, 1H), 4.13 (dd, J=11.0, 5.2 Hz, 1H), 3.91(t, J=13.4 Hz, 2H), 3.75-3.65 (m, 6H), 3.59 (dd, J=10.4, 3.2 Hz, 1H),3.43 (dd, J=6.1, 5.1 Hz, 1H), 3.15 (t, J=10.9 Hz, 1H), 2.94 (t, J=5.0Hz, 4H), 2.64 (s, 3H).

Preparation of(2R,3R,4R,5S)-5-((3-(dimethylamino)-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A339)

It was prepared according to the procedure same as that for A338. Yield:0.9 mg, 7%, white solid. LC-MS (ESI) found: 291 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 4.16 (dd, J=11.0, 5.2 Hz, 1H), 3.90 (t, J=7.7 Hz, 2H),3.76-3.67 (m, 2H), 3.59 (dd, J=10.4, 3.2 Hz, 1H), 3.45-3.41 (m, 1H),3.16 (t, J=10.9 Hz, 1H), 3.04 (s, 6H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-morpholino-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A340)

It was prepared according to the procedure same as that for A338. Yield:0.8 mg, 5%, white solid. LC-MS (ESI) found: 333 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 4.14 (dd, J=11.0, 5.2 Hz, 1H), 3.90 (t, J=12.4 Hz, 2H),3.77-3.63 (m, 6H), 3.58 (dd, J=10.4, 3.2 Hz, 1H), 3.51 (dd, J=12.3, 7.7Hz, 4H), 3.45-3.39 (m, 1H), 3.16 (t, J=10.9 Hz, 1H).

Preparation of(2R,3R,4R,5S)-5-((3-amino-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A341)

Step 1: A mixture of(2R,3R,4R,5S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A104, 30 mg, 0.11 mmol) in NH₃-MeOH solution (2.0 mL, 7 M) was stirringat 80° C. overnight. The mixture was concentrated and purified byprep-TLC to give(2R,3R,4R,5S)-5-((3-amino-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(2.1 mg, 7% yield) as a white solid. LC-MS (ESI) found: 263 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.11 (dd, J=11.1, 5.2 Hz, 1H), 3.87 (dd, J=11.0,2.6 Hz, 2H), 3.77-3.63 (m, 2H), 3.57 (dd, J=10.3, 3.3 Hz, 1H), 3.44 (dd,J=16.8, 11.7 Hz, 1H), 3.16 (t, J=10.9 Hz, 1H).

Preparation ofN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(Compound A342)

Step 1: To a mixture of(4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(200 mg, 0.46 mmol) in DCM (5 mL) was added TEA (0.13 mL, 0.923 mmol)and BzCl (98 mg, 0.69 mmol). Then the mixture was stirred reaction atroom temperature. After 2 h, the reaction was diluted with EA and water.The organic layer was separated, and concentrated in vacuo. The residuewas purified by silica gel column to giveN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(40 mg, 16% yield). LC-MS (ESI) found: 538 [M+H]⁺.

Step 2: To a solution ofN-[(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxan-3-yl]benzamide(30 mg, 0.056 mmol) in MeOH (5 mL) were added Pd/C (3 mg, 10% wt, 60%wet) and HCl (1 mL, 1 M in H₂O) at rt. The mixture was stirred at rt 2 hunder a H₂ balloon. The mixture was filtered through a Celite pad, andthe filtrate was concentrated to give productN-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide.LC-MS (ESI) found: 268 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 7.86-7.79 (m,2H), 7.56-7.48 (m, 1H), 7.45 (t, J=7.4 Hz, 2H), 4.39 (td, J=10.7, 5.3Hz, 1H), 4.04 (dd, J=11.0, 5.3 Hz, 1H), 3.91 (d, J=2.9 Hz, 1H),3.80-3.65 (m, 3H), 3.45 (dd, J=11.8, 6.5 Hz, 1H), 3.23 (t, J=10.9 Hz,1H).

The following compounds below were prepared according to the proceduresame as A342:

ID Characterization data

Yield: 21 mg, 63%, white solid. LC-MS (ESI) found: 242 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 3.98 (dd, J = 11.2, 5.3 Hz, 1H), 3.86 (d, J = 2.6 Hz,1H), 3.68 (ddd, J = 16.4, 11.4, 6.0 Hz, 2H), 3.54 (td, J = 10.6, 5.3 Hz,1H), 3.42 (dd, J = 10.4, 3.2 Hz, 1H), 3.39-3.35 (m, 1H), 3.13 (t, J =11.0 Hz, 1H), 3.03 (s, 3H).

Yield: 50 mg, 77%, colorless oil. LC-MS (ESI) found: 318 [M + Na]⁺. ¹HNMR (400 MHz, CD3OD): δ 3.97 (dd, J = 11.1, 5.3 Hz, 1H), 3.88 (d, J =2.8 Hz, 1H), 3.73-3.62 (m, 3H), 3.45 (dd, J = 10.3, 3.2 Hz, 1H),3.41-3.37 (m, 1H), 3.19 (t, J =11.1 Hz, 1H).Preparation of1-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyridin-4(1H)-one(Compound A345)

Step 1: To a solution of(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diolhydrochloride (A92, 100 mg, 0.61 mmol) and 4H-pyran-4-one (59 mg, 0.61mmol) in MeOH (2 mL) was added NaOH (24 mg, 0.61 mmol) in H₂O (1 mL).The mixture was stirred at 60° C. overnight, then purified by prep-HPLC(Method A) to give1-((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)pyridin-4(1H)-one(3.5 mg, 2% yield) as colorless oil. LC-MS (ESI) found: 242 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.08-7.56 (m, 2H), 6.61-5.93 (m, 2H), 4.26 (td,J=10.9, 5.0 Hz, 1H), 4.10 (dd, J=11.0, 5.0 Hz, 1H), 4.03 (dt, J=4.8, 3.3Hz, 2H), 3.79 (dd, J=11.4, 7.0 Hz, 1H), 3.75-3.71 (m, 1H), 3.68 (t,J=7.9 Hz, 1H), 3.63-3.59 (m, 1H).

Preparation of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carbonitrile(Compound A346)^(PGP)-9¹⁹,C³

Step 1: To a solution of(2R,3R,4R,5S)-5-((4-chloro-6-methoxypyrimidin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A303, 20 mg, 0.06 mmol) in DMF (2 mL) was added dppf (7 mg, 0.012mmol), Pd₂(dba)₃ (6 mg, 0.006 mmol) and Zn(CN)₂ (7 mg, 0.06 mmol) at rt.After the addition was complete, the mixture was stirred at 120° C.under N₂ atmosphere overnight. On consumption of starting material (LCMSmonitoring), the mixture was filtered, concentrated, and purified byflash chromatography (silica gel, 0-10% MeOH in DCM) to give4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-2-carbonitrileas a colorless oil (5 mg, 26% yield) as white solid. LC-MS (ESI) found:297 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 6.44 (s, 1H), 4.34 (td, J=10.6,5.3 Hz, 1H), 4.08 (dd, J=10.9, 5.2 Hz, 1H), 3.91 (s, 3H), 3.90 (d, J=2.7Hz, 1H), 3.72 (ddd, J=16.4, 11.4, 6.0 Hz, 2H), 3.62 (d, J=8.4 Hz, 1H),3.47-3.39 (m, 1H), 3.20-3.07 (m, 1H).

Preparation of6-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-2-methoxypyrimidine-4-carbonitrile(Compound A347)

It was prepared according to the procedure same as that for A346 byusing A327 as the starting material. LC-MS (ESI) found: 297 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.17 (s, 1H), 4.43 (dd, J=17.9, 8.3 Hz, 1H), 4.10(dd, J=13.5, 7.8 Hz, 1H), 3.96-3.85 (m, 5H), 3.75 (dd, J=11.4, 7.1 Hz,1H), 3.68 (dd, J=11.4, 5.0 Hz, 1H), 3.58 (dd, J=10.5, 3.2 Hz, 1H),3.46-3.41 (m, 1H), 3.10 (t, 1H).

Preparation of6-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)pyrimidine-2,4-dicarbonitrile(Compound A348)

It was prepared according to the procedure same as that for A346 byusing A320 as the starting material. LC-MS (ESI) found: 292 [M+H]⁺. HNMR (400 MHz, MeOD): δ7.03 (s, 1H), 4.49 (td, J=10.6, 5.1 Hz, 1H), 4.06(dd, J=11.0, 5.2 Hz, 1H), 3.91 (d, J=2.9 Hz, 1H), 3.75 (dd, J=11.4, 7.1Hz, 1H), 3.68 (dd, J=11.4, 5.0 Hz, 1H), 3.61 (dd, J=10.6, 3.2 Hz, 1H),3.48-3.39 (m, 1H), 3.12 (t, J=10.9 Hz, 1H).

Preparation of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxy-1,3,5-triazine-2-carbonitrile(Compound A349)

It was prepared according to the procedure same as that for A346 byusing A330 as the starting material. ¹H NMR (400 MHz, DMSO-d₆): δ 8.58(dd, J=26.3, 8.7 Hz, 1H), 4.19-4.12 (m, 1H), 3.90 (d, J=10.2 Hz, 3H),3.86-3.83 (m, 1H), 3.81 (d, J=5.2 Hz, 1H), 3.77 (d, J=5.2 Hz, 1H), 3.73(d, J=6.0 Hz, 2H), 3.55-3.51 (m, 1H), 3.48 (dd, J=6.1, 1.3 Hz, 2H), 3.24(dd, J=10.9, 5.5 Hz, 1H), 3.01 (td, J=10.9, 6.5 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-methoxy-6-(4-methylpiperazin-1-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A350)

Step 1: To a solution of(2R,3R,4R,5S)-5-((4-chloro-6-methoxypyrimidin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A303, 20 mg, 0.065 mmol) and DIPEA (25 mg, 0.195 mmol) in dry i-PrOH (2mL) at rt under N₂ atmosphere was added 1-methylpiperazine (17 mg, 0.13mmol). After the addition was complete, the reaction was stirred at 80°C. overnight. On consumption of starting material (TLC monitoring), thereaction vessel was again cooled to rt.

The mixture was concentrated in vacuo. The crude product was purified byflash chromatography (silica gel, 0-10% MeOH in DCM) to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-methoxy-6-(4-methylpiperazin-1-yl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diolas a yellow solid (12 mg, 50% yield). LC-MS (ESI) found: 370[M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 5.38 (s, 1H), 4.24 (td, J=10.5, 5.2 Hz, 1H), 4.16(dd, J=10.7, 5.2 Hz, 1H), 3.89 (d, J=2.7 Hz, 1H), 3.75 (dd, J=11.3, 7.0Hz, 1H), 3.68 (dd, J=11.4, 5.0 Hz, 1H), 3.63-3.57 (m, 4H), 3.56 (d,J=3.2 Hz, 1H), 3.44 (dt, J=5.9, 5.4 Hz, 1H), 3.09 (dd, J=20.7, 10.1 Hz,1H), 2.54 (t, J=5.1 Hz, 4H), 2.36 (s, 3H).

Preparation of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-(methylthio)pyrimidine-2-carbonitrile(Compound A351) and4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-(methylthio)pyrimidine-2-carboxamide(Compound A352)

Was prepared according to the procedure same as that or by using 20 asthe starting material. A351: Yield: 2.0 mg, 4%, white solid. LC-MS (ESI)found: 313 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD): δ6.46 (s, 1H), 4.41 (dd,J=12.3, 7.5 Hz, 1H), 4.05 (dd, J=12.4, 5.3 Hz, 1H), 3.89 (d, J=2.7 Hz,1H), 3.75 (dd, J=11.4, 7.1 Hz, 1H), 3.68 (dd, J=11.4, 5.0 Hz, 1H), 3.56(dd, J=10.5, 3.2 Hz, 1H), 3.47-3.40 (m, 1H), 3.10 (t, 1H), 2.49 (s, 3H).A352: Yield: 3.0 mg, 3%, white solid. LC-MS (ESI) found: 331 [M+H]⁺. ¹HNMR (400 MHz, CD₃OD): δ6.42 (s, 1H), 4.57 (dd, J=21.4, 17.8 Hz, 1H),4.03 (dd, J=10.9, 4.8 Hz, 1H), 3.91 (d, J=2.9 Hz, 1H), 3.75 (dd, J=11.3,7.1 Hz, 1H), 3.69 (dd, J=11.4, 5.0 Hz, 1H), 3.55 (dd, J=10.5, 3.0 Hz,1H), 3.45 (dd, J=12.6, 6.9 Hz, 1H), 3.13 (t, J=10.7 Hz, 1H), 2.52 (s,3H).

Preparation of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-(4-methylpiperazin-1-yl)pyrimidine-2-carbonitrile(Compound A353)

It was prepared according to the procedure same as that for A350 byusing A320 as the starting material. Yield: 2.0 mg, 3%, white solid. ¹HNMR (400 MHz, CD₃OD): δ5.79 (s, 1H), 4.21 (dd, J=21.2, 12.1 Hz, 1H),4.03 (dd, J=11.0, 5.0 Hz, 1H), 3.89 (d, J=2.9 Hz, 1H), 3.74 (dd, J=11.4,7.1 Hz, 1H), 3.67 (dd, J=11.4, 5.0 Hz, 1H), 3.61-3.51 (m, 5H), 3.45-3.39(m, 1H), 3.09 (t, J=10.9 Hz, 1H), 2.53-2.45 (m, 4H), 2.32 (s, 3H).

Preparation of(2R,3R,4R,5S)-5-(benzo[d]thiazol-2-ylamino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A354)

Step 1: A solution of(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A92, 20 mg, 0.12 mmol), 2-bromobenzo[d]thiazole (31 mg, 0.15 mmol),K₃PO₄ (78 mg, 0.37 mmol), Pd₂(dba)₃ (11 mg, 0.01 mmol) and Xantphos (14mg, 0.02 mmol) in toluene (2 mL) was stirred at 100° C. overnight. Themixture was concentrated and the residue was purified by prep-TLC togive(2R,3R,4R,5S)-5-(benzo[d]thiazol-2-ylamino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(4 mg, 9% yield) as white solid. LC-MS (ESI) found: 297 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.61-7.52 (m, 1H), 7.41 (d, J=7.6 Hz, 1H), 7.28-7.18(m, 1H), 7.10-6.88 (m, 1H), 4.25-4.06 (m, 2H), 3.91 (d, J=2.5 Hz, 1H),3.77 (dd, J=11.4, 7.0 Hz, 1H), 3.70 (dd, J=11.4, 5.0 Hz, 1H), 3.60 (dd,J=10.0, 3.2 Hz, 1H), 3.48-3.42 (m, 1H), 3.20 (d, J=12.0 Hz, 1H).

Preparation of (2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-(trifluoromethyl)phenyl) amino) tetrahydro-2H-pyran-3,4-diol (Compound A355)

It was prepared according to the procedure same as that for A354. LC-MS(ESI) found: 308 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 8.49 (s, 1H), 7.23(t, J=7.9 Hz, 1H), 6.98-6.86 (m, 2H), 6.81 (d, J=7.6 Hz, 1H), 4.03 (dd,J=11.4, 5.0 Hz, 1H), 3.91 (d, J=2.9 Hz, 2H), 3.82 (dd, J=10.4, 4.9 Hz,1H), 3.77 (dd, J=11.2, 4.3 Hz, 1H), 3.69 (dd, J=11.4, 5.0 Hz, 1H), 3.54(dd, J=10.1, 3.1 Hz, 1H), 3.47-3.40 (m, 1H), 3.05 (t, J=11.0 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyridin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A356)

Step 1: To the solution of(3S,4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxan-3-amine (100mg, 0.23 mmol) in NMP (1 mL) were added2-fluoro-4-(trifluoromethyl)pyridine (0.042 mL, 0.35 mmol) and DIPEA(0.12 mL, 0.69 mmol). The mixture was stirred at 80° C. overnight. Themixture was then concentrated and purified by flash to getN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-4-(trifluoromethyl)pyridin-2-amine(104 mg, 78% yield) as white solid. LC-MS (ESI) found: 579 [M+H]⁺.

Step 2: To the solution ofN-[(3S,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxan-3-yl]-4-(trifluoromethyl)pyridin-2-amine(80 mg, 0.14 mmol) in DCM (1 mL) was added BBr₃ (122 μL, 1 M in DCM) at−40° C. The mixture was stirred for 4 h to completion. MeOH was addedand then concentrated, the solid was filtered and washed by DCM. It waspurified by prep-TLC to get(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyridin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(7 mg, 15% yield) as white solid. LC-MS(ESI) found 309 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 8.12 (d, J=5.4 Hz, 1H), 6.78 (s, 1H), 6.71 (d, J=5.4Hz, 1H), 4.28 (td, J=10.6, 5.1 Hz, 1H), 4.11 (dd, J=11.0, 5.2 Hz, 1H),3.90 (d, J=2.7 Hz, 1H), 3.76 (dd, J=11.4, 7.1 Hz, 1H), 3.69 (dd, J=11.4,5.0 Hz, 1H), 3.58 (dd, J=10.4, 3.2 Hz, 1H), 3.45 (dd, J=12.4, 6.4 Hz,1H), 3.12 (t, J=10.9 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A357)

Step 1: A mixture of(2R,3R,4R,5S)-5—[(3-bromo-1,2,4-thiadiazol-5-yl)amino]—2-(hydroxymethyl)oxane-3,4-diol(A299, 10 mg, 0.03 mmol), 4-(tributylstannyl)pyridine (23 mg, 0.06mmol), Pd(PPh₃)₄(18 mg, 0.02 mmol) in toluene (1 ml) was stirred underN₂ at 80° C. for 12 h. The mixture was concentrated and purified byprep-HPLC (Method A) to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(0.7 mg, 2% yield) as a white solid. LC-MS (ESI) found: 325 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.62 (d, J=6.1 Hz, 2H), 8.11 (d, J=6.2 Hz, 2H),4.23 (dd, J=11.0, 5.2 Hz, 1H), 4.16-4.06 (m, 1H), 3.92 (d, J=3.0 Hz,1H), 3.77 (dd, J=11.4, 7.0 Hz, 1H), 3.70 (dd, J=11.4, 5.1 Hz, 1H), 3.65(dd, J=10.3, 3.1 Hz, 1H), 3.51-3.44 (m, 1H), 3.23 (t, J=10.9 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((3-(pyridin-3-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A358)

It was prepared according to the procedure same as that for A357. LC-MS(ESI) found: 325 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 9.26 (d, J=1.4 Hz,1H), 8.58 (dd, J=4.9, 1.6 Hz, 1H), 8.52 (dt, J=8.0, 1.9 Hz, 1H),7.54-7.49 (m, 1H), 4.23 (dd, J=11.0, 5.2 Hz, 1H), 4.12 (dd, J=10.9, 5.0Hz, 1H), 3.93 (d, J=2.8 Hz, 1H), 3.77 (dd, J=11.4, 7.1 Hz, 1H), 3.68(ddd, J=13.5, 10.8, 4.1 Hz, 2H), 3.48 (dd, J=6.5, 5.5 Hz, 1H), 3.23 (t,J=10.8 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((6-methoxy-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A359)

Step 1: To a solution of 4,6-dichloro-2-(trifluoromethyl) pyrimidine(1.0 g, 4.63 mmol) in MeOH (30 mL) was added NaOMe (750 mg, 13.89 mmol).The mixture was stirred at the room temperature for 16 h. The mixturewas diluted ethyl acetate (40 mL) and washed water (50 mL×3), theorganic layer was dried over sodium sulfate, filtered, concentratedunder reduced pressure to give 4-chloro-6-methoxy-2-(trifluoromethyl)pyrimidine. Note the purification of this reagent is missing (424 mg,43% yield). LC-MS (ESI) found: 213 [M+H]⁺.

Step 2: To a solution of4-chloro-6-methoxy-2-(trifluoromethyl)pyrimidine (42.0 mg, 0.20 mmol) iniPrOH (10 mL) was added Cs₂CO₃ (32.5 mg, 0.10 mmol), DIEA (79.0 mg, 0.60mmol),(2R,3R,4R,5S)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A92, 32.6 mg, 0.20 mmol). The mixture was stirred at 90° C. for 16 h.The mixture was concentrated under reduced pressure to give a crudeproduct, which was purified by column to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((6-methoxy-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(15 mg, 21% yield). LC-MS (ESI) found: 340 [M+H]⁺. ¹H NMR (400 MHz,MeOD) δ 6.26 (s, 1H), 4.29 (d, J=5.4 Hz, 1H), 4.07 (dd, J=10.9, 5.3 Hz,1H), 3.96-3.79 (m, 4H), 3.74-3.52 (m, 3H), 3.39 (dd, J=12.2, 6.8 Hz,1H), 3.08 (t, J=10.8 Hz, 1H).

Preparation of6-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-2-methoxypyrimidine-4-carboxamide(Compound A360)

Step 1: A stirred solution of6-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-2-methoxypyrimidine-4-carbonitrile(A346, 30 mg, 0.01 mol) in DMSO (2 mL), cooled in an ice bath, was added30% H₂O₂(0.2 mL) and K₂CO₃ (10 mg, 0.001 mol), the reaction was allowedto warm up to rt (strong exothermic effect was observed). After 5 min,distilled water (50 mL) was added, cooling applied. The mixture wasconcentrated in vacuo. The crude product was purified by pre-HPLC(Method A) to give6-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-2-methoxypyrimidine-4-carboxamideas a white solid (5.2 mg, 16% yield). LC-MS (ESI) found: 315 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.62 (s, 1H), 4.47 (s, 1H), 4.10 (s, 1H), 3.92(d, J=2.6 Hz, 4H), 3.72 (ddd, J=16.4, 11.4, 6.1 Hz, 2H), 3.62 (dd,J=10.7, 3.1 Hz, 1H), 3.51-3.39 (m, 1H), 3.13 (t, J=10.9 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((6-methoxy-2-(1H-tetrazol-5-yl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A361).

Step 1: A stirred solution of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-2-carbonitrile(A346, 30 mg, 0.01 mol) and NH₄Cl (38.3 mg, 0.3 mmol) in DMF (2 mL) atrt under N₂ atmosphere was added NaN₃ (35.8 mg, 0.2 mmol). After theaddition was complete, the reaction was stirred at 120° C. forovernight. On consumption of starting material (LCMS monitoring), thereaction vessel was again cooled to rt. The mixture was concentrated invacuo. The crude product was purified by pre-HPLC (Method A) to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((6-methoxy-2-(1H-tetrazol-5-yl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diolas a yellow solid (6.5 mg, 16% yield). LC-MS (ESI) found: 340 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.81 (s, 1H), 4.56 (s, 1H), 4.12 (s, 1H), 3.95(d, J=2.8 Hz, 4H), 3.75 (ddd, J=16.3, 11.3, 6.0 Hz, 2H), 3.65 (d, J=8.0Hz, 1H), 3.48 (dd, J=6.9, 5.3 Hz, 1H), 3.20 (t, J=10.9 Hz, 1H).Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((6-methoxy-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A362)

Step 1: To a solution of 2,4-dichloro-6-(trifluoromethyl)pyrimidine (300mg, 1.39 mmol) in MeOH (10 mL) was added NaOMe (187 mg, 3.47 mmol). Themixture was stirred at 25° C. for 16 h. The mixture was concentratedunder reduced pressure to give a crude product, which was purified bycolumn to give 2-chloro-4-methoxy-6-(trifluoromethyl)pyrimidine (200 mg,68% yield). LC-MS (ESI) found: 213 [M+H]⁺.

Step 2: To a solution of2-chloro-4-methoxy-6-(trifluoromethyl)pyrimidine (200 mg, 0.943 mmol) iniPrOH (10 mL) was added DIEA (371 mg, 2.83 mmol) and CsF (72 mg, 0.47mmol). The mixture was stirred at 90° C. for 16 h. The mixture wasconcentrated under reduced pressure to give a crude product, which waspurified by column to give(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-methoxy-6-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4—(12mg, 4% yield). LC-MS (ESI) found: 340 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ6.32 (s, 1H), 4.35 (d, J=5.1 Hz, 1H), 4.14 (dd, J=11.0, 5.3 Hz, 1H),3.95 (s, 3H), 3.90 (d, J=2.9 Hz, 1H), 3.75 (dd, J=11.4, 7.0 Hz, 1H),3.69 (t, J=5.7 Hz, 1H), 3.64 (dd, J=10.6, 3.2 Hz, 1H), 3.47-3.42 (m,1H), 3.15 (t, J=10.8 Hz, 1H)

Preparation of(2R,3R,4R,5S)-5-((2,6-dimethoxypyrimidin-4-yl)amino)-2-(hydroxy methyl)tetrahydro-2H-pyran-3,4-diol (Compound A363)

Step 1: To a solution of(3S,4S,5R,6S)-5-benzyl-4-(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(400 mg, 0.96 mmol) in DMF (20 mL) was added4-bromo-2,6-dimethoxypyrimidine (209 mg, 0.96 mmol), Ruphos.Pd.G3 (80mg, 0.10 10 mmol), Ruphos (45 mg, 0.10 mmol), Cs₂CO₃ (94 mg, 0.29 mmol).The mixture was stirred at 100° C. for 16 h under N₂. The mixture wasdiluted with water (40 mL), the organic layer was dried over sodiumsulfate, filtered, concentrated, and purified by flash column to giveN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,6-dimethoxypyrimidin-4-amine(200 mg, 37% yield). LC-MS (ESI) found: 572 [M+H]⁺.

Step 2: To a solution ofN-((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-2,6-dimethoxypyrimidin-4-amine(200 mg, 0.35 mmol) in DCM (30 mL) was added BCl₃ (0.58 mL, 1 M in DCM).The mixture was stirred at 25° C. under N₂ for 16 h. NH₃H₂O was added tothe mixture until pH was adjusted to 9. The mixture was concentratedunder reduced pressure to give a crude product, which was purified by 20column to give(2R,3R,4R,5S)-5-((2,6-dimethoxypyrimidin-4-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(16 mg, 15% yield). LC-MS (ESI) found: 302 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ: 5.47 (s, 1H), 4.18 (s, 1H), 4.04 (dd, J=11.0, 4.9 Hz, 1H),3.85 (s, 1H), 3.85 (s, 3H), 3.78 (s, 3H), 3.71 (dd, J=11.3, 7.1 Hz, 1H),3.64 (dd, J=11.4, 5.0 Hz, 1H), 3.51 (dd, J=10.4, 3.2 Hz, 1H), 3.41-3.36(m, 1H), 3.06 (t, J=10.9 Hz, 1H).

Preparation of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinonitrile(Compound A364)

Step 1: To a solution of 2-chloro-6-methoxyisonicotinic acid (400 mg,2.139 mmol) in DMF (30 mL) was added 1,1′-carbonyldiimidazole (381 mg,2.353 mmol). The mixture was stirred at 45° C. for 1 h. Then NH₃H₂O (3mL) was added to the mixture. The mixture was stirred at the roomtemperature for 2 h. The mixture was concentrated under reduced pressureto give a crude product, which was purified by column to give2-chloro-6-methoxyisonicotinamide (400 mg, 90% yield). LC-MS (ESI)found: 187 [M+H]⁺.

Step 2: To a solution of 2-chloro-6-methoxyisonicotinamide (350 mg,1.872 mmol) in DCM (50 mL) in 0° C. was added TEA (567 mg, 5.616 mmol)and TFAA (472 mg, 2.246 mmol). The mixture was stirred at the roomtemperature for 2 h. The mixture was concentrated under reduced pressureto give a crude product, which was purified by column to give2-chloro-6-methoxyisonicotinonitrile(200 mg, 64% yield). LC-MS (ESI)found: 169 [M+H]⁺.

Step 3: To a solution of 2-chloro-6-methoxyisonicotinonitrile (200 mg,1.190 mmol) in DMF (3 mL) was added Ruphos Pd G3 (99 mg, 0.119 mmol),Ruphos (57 mg, 0.119 mmol), Cs₂CO₃ (1.2 g, 3.57 mmol). The mixture wasstirred at 100° C. under N₂ for 3 h. The mixture was concentrated underreduced pressure to give a crude product, which was purified by columnto give2-(((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinonitrile(160mg, 24% yield). LC-MS (ESI) found: 566 [M+H]⁺.

Step 4: To a solution of2-(((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinonitrile(160 mg, 0.283 mmol) in DCM (10 mL) was added BCl₃ (0.566 mL, 1 M inDCM). The mixture was stirred at the room temperature for 16 h. Themixture was concentrated under reduced pressure to give a crude product,which was purified by column to give2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinonitrile(20 mg, 24% yield). LC-MS (ESI) found: 296 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 6.29 (d, J=0.9 Hz, 1H), 6.12 (d, J=0.9 Hz, 1H), 4.27 (d, J=5.6Hz, 1H), 4.15 (dd, J=11.0, 5.3 Hz, 1H), 3.90 (d, J=2.6 Hz, 1H), 3.88 (s,3H), 3.75 (dd, J=11.3, 7.1 Hz, 1H), 3.69 (dd, J=11.4, 5.1 Hz, 1H), 3.58(dd, J=10.4, 3.2 Hz, 1H), 3.47-3.42 (m, 1H), 3.09 (t, J=10.8 Hz, 1H).

Preparation of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinamide(Compound A365)

Step 1: To a solution of 2-chloro-6-methoxyisonicotinonitrile (221 mg,1.190 mmol) in DMF (5 mL) was added Ruphos Pd G3 (99 mg, 0.119 mmol),Ruphos (56 mg, 0.119 mmol), Cs₂CO₃ 15 (1.2 g, 3.57 mmol). The mixturewas stirred at 100° C. under N₂ for 3 h. The mixture was concentratedunder reduced pressure to give a crude product, which was purified bycolumn to give2-(((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinamide(80 mg, 12% yield). LC-MS (ESI) found: 584 [M+H]⁺.

Step 2: To a solution of2-(((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinamide(80 mg, 0.137 mmol) in DCM (10 mL) was added BCl₃ (0.274 mL, 1 M inDCM). The mixture was stirred at the room temperature for 16 h. Themixture was concentrated under reduced pressure to give a crude product,which was purified by column to give2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxyisonicotinonitrile(10mg, 23% yield). LC-MS (ESI) found: 314 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ6.44 (d, J=1.2 Hz, 1H), 6.28 (d, J=1.1 Hz, 1H), 4.28-4.14 (m, 2H), 3.91(d, J=2.6 Hz, 1H), 3.87 (s, 3H), 3.73 (ddd, J=16.3, 11.3, 6.0 Hz, 2H),3.59 (dd, J=10.1, 3.2 Hz, 1H), 3.45 (dd, J=6.6, 5.5 Hz, 1H), 3.11 (t,J=10.4 Hz, 1H).

Preparation of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)(methyl)amino)-6-methoxypyrimidine-2-carbonitrile(Compound A366)

Step 1: To a solution of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-2-carbonitrile(A346, 150 mg, 0.507 mmol) in DMF (10 mL) was added imidazole (69 mg,1.014 mmol) and TBDPSCI (279 mg, 1.014 mmol). The mixture was stirred at25° C. for 16 h. The mixture was concentrated under reduced pressure togive a crude product, which was purified by column to give4-(((3S,4R,5R,6R)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-4,5-dihydroxytetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-2-carbonitrile(100 mg, 37% yield). LC-MS (ESI) found: 535 [M+H]⁺.

Step 2: To a solution of4-(((3S,4R,5R,6R)-6-(((tert-butyldiphenylsilyl)oxy)methyl)-4,5-dihydroxytetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-2-carbonitrile(100 mg, 0.187 mmol) in 2,2-dimethoxypropane (10 mL) in was added TsOH(4 mg, 0.018 mmol). The mixture was stirred at the room temperature for16 h. The mixture was concentrated under reduced pressure to give acrude product, which was purified by column to4-(((3aR,4R,7S,7aR)-4-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)amino)-6-methoxypyrimidine-2-carbonitrile(70 mg, 65% yield). LC-MS (ESI) found: 575 [M+H]⁺.

Step 3: To a solution of4-(((3aR,4R,7S,7aR)-4-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)amino)-6-methoxypyrimidine-2-carbonitrile(70 mg, 0.122 mmol) in DMF (10 mL) was added NaH (9.4 mg, 0.235 mmol,60% wt. in mineral oil). The mixture was stirred at 0° C. under N₂ for 1h, then Mel (34.6 mg, 0.244 mmol) was added. The mixture was stirred atthe room temperature under N₂ for 1 h. The mixture was added drops ofsaturated aqueous NH₄C₁ and concentrated under reduced pressure to givea crude product, which was purified by column to give4-(((3aR,4R,7S,7aR)-4-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)(methyl)amino)-6-methoxypyrimidine-2-carbonitrile(25 mg, 35% yield). LC-MS (ESI) found: 589 [M+H]⁺.

Step 4: To a solution of4-(((3aR,4R,7S,7aR)-4-(((tert-butyldiphenylsilyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)(methyl)amino)-6-methoxypyrimidine-2-carbonitrile(15 mg, 0.043 mmol) in THE (1 mL) was added TBAF (0.086 mL, 1 M in THF).The mixture was stirred at the room temperature for 16 h. The mixturewas concentrated under reduced pressure to give a crude product, whichwas purified by column to give4-(((3aR,4R,7S,7aR)-4-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)(methyl)amino)-6-methoxypyrimidine-2-carbonitrile(10 mg, 67% yield). LC-MS (ESI) found: 351 [M+H]⁺.

Step 5: To a solution of4-(((3aR,4R,7S,7aR)-4-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)(methyl)amino)-6-methoxypyrimidine-2-carbonitrile(10 mg, 0.029 mmol) in THE (1 mL) was added drops of HCl (1 N in H₂O).The mixture was stirred at the room temperature for 16 h. The mixturewas concentrated under reduced pressure to give4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)(methyl)amino)-6-methoxypyrimidine-2-carbonitrile(1.5 mg, 17% yield). LC-MS (ESI) found: 311 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 6.43 (s, 1H), 3.97 (s, 2H), 3.97 (s, 3H), 3.86 (dd, J=10.8, 5.2Hz, 1H), 3.76 (dd, J=11.3, 7.1 Hz, 1H), 3.69 (dd, J=11.3, 5.0 Hz, 1H),3.49 (dd, J=6.6, 5.4 Hz, 2H), 3.13 (m, 4H).

Preparation of(2R,3R,4R,5S)-5-((4-chloro-6-ethoxypyrimidin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A367) and2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-ethoxypyrimidine-4-carbonitrile(Compound A368)

Step 1: A mixture of Na (110 mg, 4.8 mmol) in EtOH (10.0 mL) wasstirring at rt for 1 h, 2,4,6-trichloropyrimidine (0.56 mL, 4.8 mmol)was added and the mixture was further stirred for 1 h. Then the reactionmixture was concentrated and purified by chromatography on (silica gel,0-10% ethyl acetate in petroleum) to give2,4-dichloro-6-ethoxypyrimidine (630 mg, 68% yield) as a 10 white solid.LC-MS (ESI) found: 193 [M+H]⁺.

Step 2: To a mixture of 2,4-dichloro-6-ethoxypyrimidine (416 mg, 2.2mmol) in NMIP (5.0 mL) was added A92, (200 mg, 1.2 mmol) and NMN (0.71mL, 4.3 mmol) at rt under N₂. After stirring at 120° C. overnight, themixture was concentrated and purified by chromatography on (silica gel,0-50% ethyl acetate in petroleum) to give(2R,3R,4R,5S)-5-((4-chloro-6-ethoxypyrimidin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A367, 200 mg, 44% yield) as a yellow solid. LC-MS (ESI) found: 320[M+1]⁺. ¹H NMR (400 MHz, MeOD): δ 6.02 (s, 1H), 4.45-4.22 (m, 3H), 4.10(dd, J=10.8, 5.2 Hz, 1H), 3.90 (d, J=2.8 Hz, 1H), 3.77-3.58 (m, 3H),3.47-3.39 (m, 1H), 3.13 (t, J=10.8 Hz, 1H), 1.34 (t, J=7.1 Hz, 3H).

Step 3: To a mixture of(2R,3R,4R,5S)-5-((4-chloro-6-ethoxypyrimidin-2-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A367, 60 mg, 0.19 mmol) in DMF (3.0 mL) was added Zn(CN)₂ (26 mg, 0.23mmol), dppf (10.4 mg, 0.02 mmol) and Pd₂(dba)₃ (17.2 mg, 0.02 mmol) atrt under N₂. After stirring at 120° C. overnight, the reaction mixturewas concentrated and purified by prep-TLC to give2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-ethoxypyrimidine-4-carbonitrile(4.3 mg, 7% yield) as a white solid. LC-MS (ESI) found: 311 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.41 (s, 1H), 4.55-4.20 (m, 3H), 4.08 (dd,J=11.0, 5.3 Hz, 1H), 3.90 (d, J=2.9 Hz, 1H), 3.79-3.57 (m, 3H),3.48-3.39 (m, 1H), 3.13 (t, J=10.9 Hz, 1H), 1.35 (t, J=6.9 Hz, 3H).

Preparation of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-4-(trifluoromethyl)thiazole-5-carbonitrile(Compound A369)

Step 1: 2- chloro-4-(trifluoromethyl)-1,3-thiazole-5-carboxamide (200mg, 0.87 mmol) in DCM (10 ml) was added TEA (184 mg, 1.82 mmol) and TFAA(305 mg, 1.45 mmol) at 0° C., the mixture was stirred at rt for 3 h. Themixture was neutralized by NaHCO₃, extracted with DCM, purified bySilica (silica gel, 0-100% EA in PE) to give2-chloro-4-(trifluoromethyl)-1,3-15 thiazole-5-carbonitrile (70 mg, 38%yield) as solid. LC-MS (ESI) found: 212 [M+H]⁺.

Step 2: (2R,3R,4R)-5-amino-2-(hydroxymethyl)oxane-3,4-diol (A92, 50 mg,0.31 mmol), 2-chloro-4-(trifluoromethyl)-1,3-thiazole-5-carbonitrile (68mg, 0.32 mmol) and DIPEA (119 mg, 0.92 mmol) in i-PrOH (3 mL) werestirred at 120° C. for 12 h. The mixture was concentrated and purifiedby to give2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-4-(trifluoromethyl)thiazole-5-carbonitrile(18 mg, 17% yield) as solid. LC-MS (ESI) found: 315 [M+H]⁺. ¹H NMR (400MHz, CD₃OD): δ7.08 (s, 1H), 4.16 (dd, J=11.0, 5.2 Hz, 1H), 4.04 (td,J=10.5, 5.2 Hz, 1H), 3.89 (d, J=2.9 Hz, 1H), 3.74 (dd, J=11.4, 7.1 Hz,1H), 3.67 (dd, J=11.4, 5.0 Hz, 1H), 3.57 (dd, J=10.3, 3.2 Hz, 1H), 3.43(dd, J=6.5, 5.5 Hz, 1H), 3.14 (t, J=10.8 Hz, 1H).

Preparation of(2R,3R,4R,5S)-5-((6-chloro-2-(methylsulfonyl)pyrimidin-4-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A370)

Step 1: A solution of(2R,3R,4R,5S)-5-((6-chloro-2-(methylthio)pyrimidin-4-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(A331, 30 mg, 0.09 mmol), mCPBA (48 mg, 0.28 mmol) in DCM (5 mL) wasstirred at rt for 12 h. The mixture was concentrated and purified by C18column to give(2R,3R,4R,5S)-5-((6-chloro-2-(methylsulfonyl)pyrimidin-4-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(2.2 mg, 6% yield) as white solid. LC-MS (ESI) found: 354 [M+H]⁺. ¹H NMR(400 MHz, CD₃OD): δ6.65 (s, 1H), 4.49 (td, J=10.7, 5.3 Hz, 1H), 4.09(dd, J=10.9, 5.2 Hz, 1H), 3.90 (d, J=2.6 Hz, 1H), 3.75 (dd, J=11.4, 7.1Hz, 1H), 3.68 (dd, J=11.4, 5.0 Hz, 1H), 3.60 (dd, J=10.6, 3.2 Hz, 1H),3.49-3.39 (m, 2H), 3.13 (t, J=10.9 Hz, 1H).

Preparation of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-((4-(methylsulfonyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A371)

It was prepared according to the procedure same as that for A370 byusing A336 as the starting material. Yield: 8.1 mg, 25%, white solid.LC-MS (ESI) found: 320 [M+H]⁺. ¹H NMR (400 MHz, CD₃OD): δ8.59 (d, J=4.3Hz, 1H), 7.10 (d, J=4.8 Hz, 1H), 4.39 (td, J=10.2, 4.8 Hz, 1H), 4.09(dd, J=11.0, 5.2 Hz, 1H), 3.91 (d, J=2.7 Hz, 1H), 3.76 (dd, J=11.4, 7.1Hz, 1H), 3.72-3.63 (m, 2H), 3.50-3.42 (m, 1H), 3.27-3.12 (m, 4H).

Preparation of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)thiazole-5-carbonitrile(Compound A372)

It was prepared according to the procedure same as that for A363 using2-chlorothiazole-5-carbonitrile. LC-MS (ESI) found: 272 [M+H]⁺. 1H NMR(400 MHz, MeOD): δ 7.68 (s, 1H), 4.12 (dd, J=10.7, 5.2 Hz, 1H), 4.06 (m,1H), 3.89 (d, J=2.6 Hz, 1H), 3.74 (dd, J=11.4, 7.0 Hz, 1H), 3.67 (dd,J=11.4, 5.0 Hz, 1H), 3.58 (dd, J=10.1, 3.2 Hz, 1H), 3.46-3.40 (m, 1H),3.16 (t, J=10.6 Hz, 1H).

Preparation of4-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)pyrrolo[2,1-f][1,2,4]triazine-2-carbonitrile(Compound A373)

It was prepared according to the procedure same as that for A346 byusing A304 as the starting material. Yield: 6.4 mg, 13%, white solid. ¹HNMR (400 MHz, CD₃OD): δ7.63 (dd, J=2.7, 1.5 Hz, 1H), 6.99 (dd, J=4.4,1.4 Hz, 1H), 6.77 (dd, J=4.4, 2.7 Hz, 1H), 4.73 (td, J=10.7, 5.2 Hz,1H), 4.11 (dd, J=11.0, 5.3 Hz, 1H), 3.94 (d, J=2.7 Hz, 1H), 3.78 (dd,J=6.2, 5.2 Hz, 1H), 3.75 (d, J=3.5 Hz, 1H), 3.70 (dd, J=11.4, 5.0 Hz,1H), 3.50-3.44 (m, 1H), 3.25 (t, J=10.9 Hz, 1H).

Preparation of methyl2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carboxylate(Compound A374)

To a stirred solution of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carbonitrile(A346, 30 mg, 0.01 mol) in MeOH (5 mL) at rt under N₂ atmosphere wasadded CH₃ONa (0.1 mL, 5 M in MeOH). After the addition was complete, thereaction was stirred at rt overnight. On consumption of startingmaterial (LCMS monitoring), the mixture was adjusted pH to 5 andconcentrated in vacuo. The crude product was purified by pre-HPLC(Method A) to give methyl2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carboxylateas a white solid (1.5 mg, 5% yield). LC-MS (ESI) found: 330[M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.62 (s, 1H), 4.36 (d, J=5.8 Hz, 1H), 4.13 (s,1H), 3.93 (d, J=23.1 Hz, 4H), 3.72 (ddd, J=16.3, 11.4, 6.1 Hz, 2H),3.65-3.59 (m, 1H), 3.46 (dd, J=11.5, 5.3 Hz, 1H), 3.31 (dd, J=3.1, 1.5Hz, 3H), 3.16 (dd, J=18.2, 7.3 Hz, 1H).

Preparation of methyl2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carbimidate(Compound A375)

To a stirred solution of2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carbonitrile(A346, 30 mg, 0.01 mol) in MeOH (5 mL) at rt under N₂ atmosphere wasadded CH₃ONa (36 mg, 0.2 mmol). After the addition was complete, thereaction was stirred at rt overnight. On consumption of startingmaterial (LCMS monitoring), the mixture was concentrated in vacuo. Thecrude product was directly purified by pre-HPLC under basic condition togive methyl2-(((3S,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)amino)-6-methoxypyrimidine-4-carboxylateas a white solid (1.5 mg, 5% yield). LC-MS (ESI) found: 329[M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 6.43 (s, 1H), 4.42 (s, 1H), 4.13 (dd, J=11.0, 5.3Hz, 1H), 3.94 (d, J=23.7 Hz, 4H), 3.73 (ddd, J=16.4, 11.4, 6.1 Hz, 2H),3.63 (dd, J=10.6, 3.2 Hz, 1H), 3.51-3.40 (m, 1H), 3.36-3.30 (m, 3H),3.14 (dd, J=12.8, 9.0 Hz, 1H).

1—OMe-α-galactose seriesPreparation of(2R,3R,4R,5R,6S)-5-((5-chloro-3-fluoropyridin-2-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A376)

Step 1: To a mixture of(2R,3R,4R,5R,6S)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A91, 50 mg, 0.25 mmol) in NMIP (2 mL) were added5-chloro-2,3-difluoropyridine (116 mg, 0.75 mmol) and DIEA (0.17 mL, 1.0mmol) at rt under N₂. After stirring at 120° C. overnight, the mixturewas concentrated and purified by prep-HPLC (Method A) to give(2R,3R,4R,5R,6S)-5-((5-chloro-3-fluoropyridin-2-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(3.9 mg, 7.8% yield) as a white solid. No LCMS. ¹H NMR (400 MHz, MeOD):δ 8.51 (s, 1H), 7.79 (d, J=2.1 Hz, 1H), 7.37 (dd, J=10.7, 2.1 Hz, 1H),4.83 (s, 1H), 4.56 (d, J=3.7 Hz, 1H), 3.93 (d, J=3.1 Hz, 1H), 3.86 (dd,J=10.8, 3.2 Hz, 1H), 3.82-3.79 (m, 1H), 3.77-3.71 (m, 2H), 3.37 (s, 3H).

The following compounds below were prepared according to the proceduresame as A376:

ID Characterization data Starting Material

Yield: 4.1 mg, 7.8%, white solid. LC-MS (ESI) found: 340 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.51 (d, J = 4.8 Hz, 1H), 6.90 (d, J = 4.9 Hz,1H), 4.81 (s, 1H), 4.52 (s, 1H), 3.93 (d, J = 2.8 Hz, 1H), 3.88-3.79 (m,2H), 3.78-3.70 (m, 2H), 3.38 (s, 3H).

Yield: 0.8 mg, 1.9%, white solid. LC-MS (ESI) found: 278 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 7.82 (s, 1H), 4.83-4.82 (m, 1H), 4.17 (t, J = 4.9Hz, 1H), 3.91 (d, J = 2.9 Hz, 1H), 3.85- 3.79 (m, 2H), 3.76-3.73 (m,2H), 3.40 (s, 3H).

Yield: 1.2 mg, 1.4%, white solid. LC-MS (ESI) found: 336 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.82 (d, J = 3.7 Hz, 1H), 4.26 (s, 1H), 3.90 (d,J = 3.1 Hz, 1H), 3.80 (dd, J = 10.6, 3.0 Hz, 2H), 3.73 (dd, J = 6.9, 5.9Hz, 2H), 3.39 (s, 3H).

Yield: 3.2 mg, 7.2%, white solid. LC-MS (ESI) found: 289 [M + H]⁺. ¹HNMR(400 MHz, MeOD): δ 7.46 (dd, J = 16.3, 8.1 Hz, 1H), 6.43 (dd, J =8.1, 2.2 Hz, 1H), 6.06 (dd, J = 7.6, 1.9 Hz, 1H), 4.79 (d, J = 3.6 Hz,1H), 4.33 (dd, J = 10.8, 3.6 Hz, 1H), 3.91 (d, J = 3.0 Hz, 1H),3.83-3.72 (m, 4H), 3.37 (s, 3H).

Yield: 8 mg, 8.6%, white solid. LC-MS (ESI) found: 339 [M + H]⁺. ¹HNMR(400 MHz, MeOD): δ 7.53 (t, J = 7.9 Hz, 1H), 6.88 (d, J = 7.2 Hz,1H), 6.80 (d, J = 8.6 Hz, 1H), 4.87 (s, 1H), 4.44 (dd, J = 10.8, 3.6 Hz,1H), 3.92 (d, J = 3.0 Hz, 1H), 3.86- 3.71 (m, 4H), 3.37 (s, 3H)

Preparation of(2R,3R,4R,5R,6S)-5-((3-(dimethylamino)-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A114)

A solution of(2R,3R,4R,5R,6S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(10 mg, 0.03 mmol), dimethylamine (2.2 mg, 0.05 mmol) and DIEA (11.6 mg,0.09 mmol) in i-PrOH (4 mL) was stirred at 120° C. overnight. Themixture was concentrated in vacuo. The crude product was purified byprep-HPLC (Method A) to give(2R,3R,4R,5R,6S)-5-((3-(dimethylamino)-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(0.8 mg, 8% yield) as white solid. LC-MS (ESI) found: 321 [M+H]⁺. ¹H NMR(400 MHz, Methanol-d₄): δ 4.85-4.81 (m, 1H), 4.12 (d, J=13.7 Hz, 1H),3.90 (d, J=3.2 Hz, 1H), 3.81 (ddd, J=11.4, 7.8, 3.3 Hz, 2H), 3.77-3.67(m, 2H), 3.39 (s, 3H), 3.04 (s, 6H).

The following compound below was prepared according to the proceduresame as A114.

Yield: 0.5 mg, 0.1%, white solid. LC- MS (ESI) found: 363 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.78 (d, J = 9.0 Hz, 1H), 4.21-4.09 (m, 1H), 3.90(d, J = 2.4 Hz, 1H), 3.85-3.76 (m, 2H), 3.76-3.67 (m, 6H), 3.56-3.46 (m,4H), 3.39 (s, 3H).1-OMe-β-galactose seriesPreparation of(2R,3R,4R,5R,6R)-5-((3,5-difluoropyridin-2-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A383)

A solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(30 mg, 0.16 mmol), DIPEA (91 mg, 0.70 mmol) and 2,3,5-trifluoropyridine(93 mg, 0.70 mmol) in i-PrOH (2 mL) was stirred at 120° C. overnight.The mixture was concentrated and the residue was purified by prep-HPLC(Method A) to give(2R,3R,4R,5R,6R)-5-((3,5-difluoropyridin-2-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(1.2 mg, 2% yield) as a white solid. LC-MS (ESI) found: 307 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 7.12 (dd, J=4.7, 2.5 Hz, 2H), 4.25 (d, J=8.1 Hz,1H), 3.88 (d, J=3.0 Hz, 1H), 3.81-3.75 (m, 2H), 3.65 (dd, J=10.5, 3.5Hz, 2H), 3.53 (t, J=6.1 Hz, 1H), 3.45 (s, 3H).

The following compounds below were prepared according to the proceduresame as A383:

Starting ID Characterization data Material

Yield: 1.2 mg, 2%, white solid. LC-MS (ESI) found: 307 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.12 (dd, J = 4.7, 2.5 Hz, 2H), 4.25 (d, J = 8.1 Hz,1H), 3.88 (d, J = 3.0 Hz, 1H), 3.81-3.75 (m, 2H), 3.65 (dd, J = 10.5,3.5 Hz, 2H), 3.53 (t, J = 6.1 Hz, 1H), 3.45 (s, 3H).

Yield: 12.1 mg, 14%, white solid. LC-MS (ESI) found: 340 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.48 (d, J = 4.9 Hz, 1H), 6.86 (d, J = 4.9 Hz,1H), 4.42 (d, J = 8.4 Hz, 1H), 4.32-4.23 (m, 1H), 3.88 (d, J = 3.0 Hz,1H), 3.76 (dt, J = 12.4, 7.1 Hz, 3H), 3.54 (t, J = 6.1 Hz, 1H), 3.44 (s,3H).

Yield: 32.3 mg, 33%, white solid. LC-MS (ESI) found: 321 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.22 (m, 1H), 8.18(s, 1H), 4.71-4.59 (m, 2H),4.58-4.50 (m, 1H), 4.44-4.33 (dd, J = 11.4, 7.2 Hz, 1H), 4.04-3.96 (d,1H), 3.78-3.63(m, 3H), 3.61-3.55 (m, 2H), 3.30 (s, 3H).

Yield: 30.4 mg, 39.0%, colorless oil. LC-MS (ESI) found: 302[M + H]⁺. ¹HNMR (400 MHz, D₂O): δ 7.52 (s, 1H), 4.39 (d, J = 8.2 Hz, 1H), 3.92 (d, J= 3.0 Hz, 1H), 3.74 (m, 3H), 3.64 (m, 2H), 3.45 (s, 3H).

Preparation of(2R,3R,4R,5R,6R)-5-((1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compouund A110)

Step 1: A solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(A90, 30 mg, 0.16 mmol), DIPEA (40 mg, 0.31 mmol) and5-chloro-1,2,4-thiadiazole (22.4 mg, 0.19 mmol) in i-PrOH (1 mL) wasstirred at 120° C. overnight. The mixture was concentrated and theresidue was purified by prep-HPLC (Method A) to give 10(2R,3R,4R,5R,6R)-5-((1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(6.8 mg, 16% yield) as a white solid. LC-MS (ESI) found: 277 [M+H]⁺. ¹HNMR (400 MHz, CD₃OD): δ7.82 (s, 1H), 4.34 (d, J=8.1 Hz, 1H), 3.87 (d,J=3.1 Hz, 1H), 3.83-3.68 (m, 4H), 3.55-3.51 (m, 1H), 3.47 (s, 3H).

Preparation of6-(((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)amino)-1,3,5-triazine-2,4(1H,3H)-dione(Compound A112)

Step 1: To a solution of 2,4,6-trichloro-1,3,5-triazine (187 mg 1.03mmol) in THE (5 mL) was added DIPEA (200 mg, 1.55 mmol) at −78° C.(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(100 mg, 0.51 mmol) was then added at −78° C. The mixture was furtherstirred for 2 h at −78° C. Then it was quenched by adding H₂O (5 mL).The mixture was warmed to rt and stirred for another 2 h. The solventwas evaporated and the residual was purified by prep-HPLC (Method A) togive6-(((2R,3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-2-methoxytetrahydro-2H-pyran-3-yl)amino)-1,3,5-triazine-2,4(1H,3H)-dione(6.1 mg, 4% yield) as white solid. LC-MS (ESI) found: 305 [M+H]⁺. ¹H NMR(400 MHz, D₂O): 6 4.39 (d, J=8.4 Hz, 1H), 4.02 (dd, J=10.7, 8.5 Hz, 1H),3.88 (d, J=3.2 Hz, 1H), 3.81-3.67 (m, 3H), 3.63 (dd, J=7.8, 4.3 Hz, 1H).

Preparation of(2R,3R,4R,5R,6S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol (Compound A105) and(2R,3R,4R,5R,6R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(Compound A106)

Step 1: A solution of(2R,3R,4R,5R,6R)-5-amino-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(224.0 mg, 1.2 mmol), 3,5-dichloro-1,2,4-thiadiazole (372.0 mg, 2.4mmol) and DIEA (464.4 mg, 3.6 mmol) in i-PrOH (10 mL) was stirred at rtovernight. The mixture was concentrated in vacuo. The crude product waspurified by flash chromatography (silica gel, 0-10% MeOH in DCM) andpre-HPLC (Method B) to give(2R,3R,4R,5R,6S)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(0.8 mg, 0.2% yield) as a white solid and(2R,3R,4R,5R,6R)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)-6-methoxytetrahydro-2H-pyran-3,4-diol(3.7 mg, 1% yield) as white solid. A105: LC-MS (ESI) found: 312 [M+H]⁺.¹H NMR (400 MHz, Methanol-d₄): δ 4.85-4.80 (m, 1H), 4.23-4.11 (m, 1H),3.91 (d, J=3.1 Hz, 1H), 3.85-3.77 (m, 2H), 3.76-3.69 (m, 2H), 3.40 (s,3H). A106: LC-MS (ESI) found: 312 [M+H]⁺. ¹H NMR (400 MHz, Methanol-d₄):δ 4.33 (d, J=8.1 Hz, 1H), 3.87 (d, J=3.2 Hz, 1H), 3.84-3.70 (m, 2H),3.68 (dd, J=10.4, 3.3 Hz, 1H), 3.53 (ddd, J=6.7, 5.4, 1.1 Hz, 1H), 3.48(s, 3H), 3.46-3.40 (m, 1H).

Talose Series

Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A387).

Step 1: A solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(50 mg, 0.12 mmol), 2-chloro-4-(trifluoromethyl)pyrimidine (32 mg, 0.1720 mmol) and DIPEA (30 mg, 0.23 mmol) in i-PrOH (2 mL) was stirred at80° C. overnight. The mixture was concentrated and the residue waspurified by flash to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-4-(trifluoromethyl)pyrimidin-2-amine(50 mg, 75% yield) as a colorless oil. LC-MS (ESI) found: 580 [M+H]⁺.

Step 2: To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-4-(trifluoromethyl)pyrimidin-2-amine(25 mg, 0.043 mmol) in DCE (2 mL) was added BCl₃ (0.43 mL, 1 M in THF)at −78° C. After stirring at-78° C. for 1 h, the mixture was quenchedwith MeOH. The mixture was concentrated and the residue was purified byprep-TLC (Method A) to give(2R,3R,4R,5R)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(2.7 mg, 20% yield) as a colorless oil. LC-MS (ESI) found: 310 [M+H]⁺.¹H NMR (400 MHz, MeOD): δ 8.50 (s, 1H), 6.89 (d, J=4.9 Hz, 1H), 4.32 (d,J=3.2 Hz, 1H), 4.02 (dd, J=12.0, 1.9 Hz, 1H), 3.95-3.92 (m, 1H),3.86-3.76 (m, 2H), 3.70 (dd, J=11.5, 4.8 Hz, 1H), 3.58 (dd, J=12.0, 1.6Hz, 1H), 3.44 (ddd, J=6.9, 4.8, 1.2 Hz, 1H).

The following compounds below were prepared according to the proceduresame as A387:

Starting ID Characterization data Material

Yield: 13.4 mg, 59%, white solid. LC-MS(ESI) found 293 [M + H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.77 (d, J = 2.1 Hz, 1H), 7.34 (dd, J = 10.7, 2.1 Hz,1H), 4.35 (d, J = 3.4 Hz, 1H), 4.02 (dd, J = 11.9, 1.9 Hz, 1H),3.94-3.91 (m, 1H), 3.81 (ddd, J = 18.5, 8.0, 5.1 Hz, 2H), 3.69 (dd, J =11.5, 4.8 Hz, 1H), 3.55 (dd, J = 12.0, 1.6 Hz, 1H), 3.47-3.41 (m, 1H).

Yield: 5.0 mg, 37%, colorless oil. LC-MS (ESI) found: 314 [M + H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.26 (d, J = 6.1 Hz, 1H), 3.99-3.89 (m, 2H), 3.78(ddd, J = 10.3, 9.1, 5.6 Hz, 3H), 3.68 (dd, J = 11.5, 4.6 Hz, 1H), 3.60(d, J = 12.3 Hz, 1H), 3.50-3.37 (m, 3H), 2.19 (d, J = 5.1 Hz, 2H).

Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(Compound A136)

Step 1: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-amine(80 mg, 0.19 mmol) and TEA (0.03 mL, 0.19 mmol) in dry DCM(3 mL) at 0°C. under N₂ atmosphere was added dropwise benzoyl chloride (27.1 mg,0.19 mmol). The reaction mixture was stirred for 1 h. The resultingmixture was diluted with DCM (30 mL), washed with H₂O (20 mL×2) andbrine (30 mL), dried over Na₂SO₄. The organic layer was separated andconcentrated in vacuo. The crude product was purified by flashchromatography (silica gel, 0˜80% EA in PE) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(70 mg, 71% yield) as a colorless oil. LC-MS (ESI) found: 538 [M+H]⁺.

Step 2: To a solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)benzamide(35 mg, 0.07 mmol) in MeOH (3 mL) was added Pd/C (5 mg, 10% wt, 60% wet)and HCl (1 mL, 1 M in H₂O) at rt under a H₂ balloon. The reaction wasstirred at rt for 3 h. The resulting mixture was filtered andconcentrated in vacuo. The crude product was purified by prep-HPLC(Method A) to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)benzamide(9.6 mg, 55%) as a colorless oil. LC-MS (ESI) found: 268 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 7.86-7.80 (m, 2H), 7.58-7.51 (m, 1H), 7.49-7.44 (m,2H), 4.31-4.27 (m, 1H), 4.04 (dd, J=12.0, 1.9 Hz, 1H), 3.98-3.95 (m,1H), 3.86 (dd, J=4.4, 3.2 Hz, 1H), 3.80 (dd, J=11.4, 7.0 Hz, 1H), 3.70(dd, J=11.5, 4.8 Hz, 1H), 3.62 (dd, J=12.0, 1.6 Hz, 1H), 3.46 (ddd,J=6.9, 4.8, 1.1 Hz, 1H).

Additional ASGPR Ligands

Preparation ofN-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(Compound A183)

Step 1: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-carbaldehyde(1.2 g, 2.7 mmol) in dry MeOH (10 mL) at 0° C. under N₂ atmosphere wasadded NaBH₄ (230 mg, 6.05 mmol) in portions. After the addition wascomplete, the reaction was stirred at rt overnight. On consumption ofstarting material (TLC monitoring), the reaction vessel was again cooledto 0° C. The reaction mixture was quenched with 1 mL ice water. Themixture was concentrated in vacuo. The crude product was purified byflash chromatography (silica gel, 0-20% MeOH in DCM) to give((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanol(1 g, 83% yield) as a colorless oil. LC-MS (ESI) found: 447 [M+H]⁺. ¹HNMR (400 MHz, CDCl₃): δ 7.37-7.26 (m, 15H), 6.39 (s, 1H), 4.82 (dd,J=11.7, 7.4 Hz, 2H), 4.68-4.40 (m,4H), 4.33 (d, J=3.7 Hz, 1H), 4.21 (t,J=4.5 Hz, 1H), 4.02 (ddd, J=20.7, 16.4, 11.9 Hz, 3H), 3.74 (ddd, J=15.5,10.2, 6.2 Hz, 2H).

Step 2: To a solution of((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanol(1 g, 2.24 mmol) in MeOH (40 mL) was added Pd/C (100 mg, 10% wt, 60%wet). The reaction mixture was stirred at rt under a H₂ balloon for 2 h.The mixture was filtered and concentrated in vacuo. The crude productwas purified by flash chromatography (silica gel, 0-20% MeOH in DCM) togive((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanol(350 mg, 35% yield) as a colorless oil. LC-MS (ESI) found: 449 [M+H]⁺.¹H NMR (400 MHz, CDCl₃): δ 7.38-7.25 (m, 15H), 4.88 (dd, J=13.5, 11.6Hz, 1H), 4.69-4.50 (m, 4H), 4.44 (dd, J=11.9, 8.1 Hz, 1H), 4.01 (d,J=5.0 Hz, 1H), 3.87-3.72 (m, 3H), 3.63-3.45 (m, 3H), 2.04 (dd, J=7.0,4.0 Hz, 1H), 1.17 (d, J=7.1 Hz, 2H).

Step 3: To a solution of((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanol(350 mg, 0.78 mmol) and TEA (394 mg, 3.90 mmol) in DCM (10 mL) was addedTsCl (446 mg, 2.34 mmol) slowly at 0° C. The reaction mixture wasstirred at rt overnight. The resulting mixture was extracted with EA (50mL), washed with H₂O (40 mL×2) and brine (40 mL), dried over Na₂SO₄,filtered. The organic layer was separated and concentrated in vacuo. Thecrude product was purified by flash chromatography (silica gel, 0-50% EAin PE) to give((4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl4-methylbenzenesulfonate (180 mg, 38% yield) as a yellow oil. LC-MS(ESI) found: 603 [M+H]⁺.

Step 4: To a solution of(2R,3R,4R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyran(185 mg, 0.31 mmol) in dry DMF (10 mL) was added NaN₃ (180 mg, 3.10mmol). The reaction mixture was stirred at 80° C. overnight. The mixturewas extracted with EA (20 mL), washed with H₂O (20 mL×2) and brine (20mL), dried over Na₂SO₄, filtered. The organic layer was separated andconcentrated in vacuo. The crude product was purified by flashchromatography (silica gel, 0˜50% EA in PE) to give(2R,3R,4R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyran(65 mg, 45% yield) as a colorless oil. LC-MS (ESI) found: 496 [M+Na]⁺.

Step 5: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(65 mg, 0.14 mmol) in THE (5 mL) was added PPh₃ (72 mg, 0.27 mmol) andwater (5 mL). The reaction mixture was stirred at rt overnight. Themixture was concentrated in vacuo. The crude product was purified byflash chromatography (silica gel, 0-10% MeOH in DCM) to give((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(60 mg, 98% yield) as a colorless oil. LC-MS (ESI) found: 448 [M+H]⁺.

Step 6: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(30 mg, 0.067 mmol) in dry DCM (5 mL) was added DIPEA (30 mg, 0.07 mmol)and acetyl chloride (11 mg, 0.14 mmol) dropwise at 0° C. under N₂atmosphere. The reaction mixture was stirred at rt for 8 h. The reactionwas cooled to 0° C. and quenched with saturated sodium bicarbonatesolution. The mixture was extracted with EA (10 mL), washed with H₂O (10mL×2) and brine (10 mL), dried over Na₂SO₄, filtered. The organic layerwas separated and concentrated in vacuo. The crude product was purifiedby flash chromatography (silica gel, 0˜50% EA in PE) to giveN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(14 mg, 44% yield) as a colorless oil. LC-MS (ESI) found: 490 [M+H]⁺.

Step 7: To a solution ofN-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(14 mg, 0.03 mmol) in dry MeOH (5 mL) was added Pd/C (10 mg, 10% wt, 60%wet). The reaction mixture was charged with H₂ and stirred at rt for 3days with a H₂ balloon. The mixture was filtered and concentrated invacuo to giveN-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)acetamide(1 mg, 16% yield) as a colorless oil. LC-MS (ESI) found: 220 [M+H]⁺. ¹HNMR (400 MHz, CD₃OD): δ3.91 (dd, J=11.9, 2.8 Hz, 1H), 3.85-3.72 (m, 3H),3.66 (dd, J=11.7, 4.3 Hz, 1H), 3.58 (dd, J=13.9, 4.3 Hz, 1H), 3.48-3.40(m, 3H), 1.93 (s, 3H), 1.92-1.84 (m, 1H).

General Procedure for the Mitsunobu Reaction

To a solution of((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)methanol(A182-2, 100 mg, 0.224 mmol, 1.0 eq), PPh₃ (88 mg, 0.336 mmol, 1.5 eq)and nucleophiles (1.0 eq) in dry DCM (1.1 mL) was added DIAD (0.053 mL,0.269 mmol, 1.2 eq) dropwise at ice-bath under N₂ atmosphere. Then thereaction was allowed to warm to rt. The resulting reaction mixture wasstirred at the same temperature for another 40 min, at which time TLCshowed the disappearance of all starting material. The mixture wasevaporated. The crude product was further purified by silica gel columnchromatography to give desired products.

General Procedure for the Hydrogenation Reaction

A suspension of substrate (1.0 eq) and Pd/C (0.2 eq, 10% wt, 60% wet) inMeOH was charged with H₂ and stirred under a H₂ balloon. The reactionwas stirred at rt and monitored by TLC. When TLC showed thedisappearance of all starting material, the mixture was filtered andevaporated. The crude product was further purified by silica gel columnchromatography to give desired products.Preparation of(2R,3R,4R,5R,6S)-2-(hydroxymethyl)-5-methyl-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A192),(2R,3R,4R,5S,6S)-2-(hydroxymethyl)-5-methyl-6-phenoxytetrahydro-2H-pyran-3,4-diol(Compound A191) and(2R,3R,4R,5S)-2-(hydroxymethyl)-5-(phenoxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A193)

Step 1: They were synthesized according to the general procedure. Yieldfor(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methylene-6-phenoxytetrahydro-2H-pyran:30 mg, 26%. LC-MS (ESI) found: 545 [M+Na]⁺. Yield for(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(phenoxymethyl)-3,4-dihydro-2H-pyran:70 mg, 60%. LC-MS (ESI) found: 545 [M+Na]⁺.

Step 2: They were synthesized from 100 mg of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-methylene-6-phenoxytetrahydro-2H-pyranaccording to the general procedure. Yield (A192): 0.5 mg, 1%. Yield(A191): 7 mg, 14%. LC-MS (ESI) found: 277 [M+Na]⁺. ¹H NMR (400 MHz,CD₃OD) 6 7.30-7.22 (m, 2H), 7.13-7.06 (m, 2H), 6.96 (tt, J=7.4, 1.1 Hz,1H), 5.44 (d, J=1.8 Hz, 1H), 4.11 (dd, J=5.5, 3.5 Hz, 1H), 3.89 (d,J=2.2 Hz, 2H), 3.75-3.68 (m, 2H), 2.20 (ddd, J=7.4, 5.3, 1.8 Hz, 1H),1.23 (d, J=7.4 Hz, 3H).

Step 3: 9.8 mg of(2R,3R,4R,5S)-2-(hydroxymethyl)-5-(phenoxymethyl)tetrahydro-2H-pyran-3,4-diolwas obtained from 100 mg of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-(phenoxymethyl)-3,4-dihydro-2H-pyranaccording to the general procedure. Yield: 20%. LC-MS (ESI) found: 277[M+Na]⁺. ¹H NMR (400 MHz, CD₃OD): δ7.30-7.19 (m, 2H), 6.95-6.84 (m, 3H),4.37 (t, J=10.0 Hz, 1H), 4.24 (ddd, J=9.7, 3.0, 1.4 Hz, 1H), 4.14 (dd,J=11.7, 1.9 Hz, 1H), 3.91 (dd, J=5.6, 3.2 Hz, 1H), 3.81-3.74 (m, 2H),3.66 (dd, J=11.5, 4.6 Hz, 1H), 3.51 (ddd, J=11.7, 2.6, 1.4 Hz, 1H), 3.41(ddd, J=7.2, 4.6, 1.7 Hz, 1H), 2.22 (ddd, J=10.2, 5.4, 2.7 Hz, 1H).

Synthesis 5-38. Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(3-hydroxypropyl)tetrahydro-2H-pyran-3,4-diol(Compound A162)

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(A122-1, 100 mg, 0.18 mmol) in dioxane (5 mL) under N₂ were addedPd(PPh₃)₂C1 ₂ (12.9 mg, 0.018 mmol), CuI (3.5 mg,0. 018 mmol), TEA (0.03mL,0.55 mmol). The mixture was stirred at 100° C. under N₂ for 16 h. Themixture was filtered, the filtrate was concentrated under reducedpressure to give crude product, which was purified by flashchromatography (silica gel, 10-50% EtOAc in PE) to give3-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)prop-2-yn-1-ol(30 mg, 35% yield) as colorless oil. LC-MS (ESI) found: 493 [M+Na]⁺.

Step 2: To a solution of3-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)prop-2-yn-1-ol(20 mg, 0.043 mmol) in MeOH (10 mL) was added Pd/C (5 mg, 10% wt, 60%wet), the mixture was stirred at rt under a H₂ balloon for 12 h. Themixture was filtered, the filtrate was concentrated to give a crudeproduct, which was purified by prep-HPLC (Method A) to give(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(3-hydroxypropyl)tetrahydro-2H-pyran-3,4-diol(5 mg, 57% yield). ¹H NMR (400 MHz, MeOD): δ 3.96-3.73 (m, 3H), 3.67(dd, J=11.8, 3.9 Hz, 1H), 3.54 (dt, J=7.4, 5.1 Hz, 2H), 3.42 (dd,J=11.8, 3.2 Hz, 1H), 1.73-1.41 (m, 4H), 1.40-1.29 (m, 2H), 0.91 (dd,J=14.5, 7.2 Hz, 1H).

Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-((4-methylpiperazin-1-yl)methyl)tetrahydro-2H-pyran-3,4-diol (Compound A390)

It was prepared according to the procedure same as that for A209. LC-MS(ESI) found: 261 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 4.06-3.54 (m, 16H),3.46-3.41 (m, 1H), 3.03 (s, 3H), 2.51-2.42 (m, 1H).

Preparation of((2R,3R,4R,5R)-2-(hydroxymethyl)-5-(piperidin-1-yl)tetrahydro-2H-pyran-3,4-diol(Compound A391)

It was prepared according to the procedure same as that for A222. LC-MS(ESI) found: 232 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 4.47 (dd, J=14.3, 1.6Hz, 1H), 4.10 (dd, J=10.9, 7.9 Hz, 2H), 4.03-3.93 (m, 2H), 3.75 (dd,J=11.4, 7.0 Hz, 1H), 3.71-3.65 (m, 2H), 3.61-3.53 (m, 2H), 3.26-3.18 (m,2H), 2.07 (d, J=12.7 Hz, 1H), 2.02-1.93 (m, 1H), 1.86-1.76 (m, 2H),1.73-1.56 (m, 2H).

Preparation of1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-3-methylimidazolidin-2-one(Compound A392)

Step 1: To a solution of(2R,3S,4S)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-5-iodo-3,4-dihydro-2H-pyran(200 mg, 0.369 mmol) in DMF (10 mL) was added 1-methylimidazolidin-2-one(37 mg, 0.37 mmol), CuI (7 mg, 0.37 mmol), Cs₂CO₃ (360 mg, 1.12 mmol).The mixture was stirred at 100° C. for 16 h. The mixture wasconcentrated under reduced pressure to give a crude product, which waspurified by column to give1-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)-3-methylimidazolidin-2-one(103 mg). LC-MS (ESI) found: 515 [M+H]⁺.

Step 2: To a solution of1-((2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran-5-yl)-3-methylimidazolidin-2-one(103 mg, 0.20 mmol) in MeOH (10 mL) was added Pd/C (10 mg, 10% wt, 60%wet). The mixture was stirred at the room temperature at rt under a H₂balloon for 16 h. The mixture was filtered, the filtrate wasconcentrated under reduced pressure to give1-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-3-methylimidazolidin-2-one(25 mg, 50% yield). LC-MS (ESI) found: 247 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 4.17-4.04 (m, 2H), 3.99 (s, 1H), 3.89-3.77 (m, 3H), 3.72-3.62(m, 2H), 3.47 (dd, J=11.4, 7.2 Hz, 2H), 3.38-3.33 (m, 1H), 3.26 (d,J=8.5 Hz, 1H), 2.76 (s, 3H).

Preparation of(2R,3R,4R,5R)-5-(dimethylamino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A393)

It was prepared according to the procedure same as that for A222. LC-MS(ESI) found: 192 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 4.42 (dd, J=14.3, 2.0Hz, 1H), 4.08-4.01 (m, 1H), 3.98-3.94 (m, 1H), 3.77 (dd, J=11.5, 7.0 Hz,1H), 3.66 (ddd, J=14.3, 11.9, 3.1 Hz, 2H), 3.55-3.51 (m, 1H), 3.42 (d,J=1.8 Hz, 1H), 3.13 (s, 3H), 3.01 (s, 3H).

Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-3,4-diol(Compound A394)

It was prepared according to the procedure same as that for A164. LC-MS(ESI) found: 229 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 7.58 (s, 1H), 7.48(s, 1H), 4.06 (dd, J=11.6, 5.1 Hz, 1H), 3.96 (dd, J=11.8, 7.7 Hz, 1H),3.89 (dd, J=4.6, 3.5 Hz, 1H), 3.86-3.80 (m, 4H), 3.71 (dd, J=11.9, 3.9Hz, 1H), 3.65 (m, 2H), 2.95 (dd, J=8.7, 4.6 Hz, 1H).

Preparation of(2R,3R,4R,5R)-2-(hydroxymethyl)-5-(pyrimidin-5-yl)tetrahydro-2H-pyran-3,4-diol(Compound A395)

It was prepared according to the procedure same as that for A164. LC-MS(ESI) found: 227[M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 8.98 (s, 2H), 8.92 (s,1H), 4.36 (dd, J=12.3, 2.1 Hz, 1H), 4.01 (dd, J=6.0, 3.2 Hz, 1H),3.94-3.80 (m, 3H), 3.71 (dd, J=11.6, 4.3 Hz, 1H), 3.59 (m, 1H), 3.00 (m,1H).

Preparation ofN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-N-methylacetamide(Compound A396)

Step 1: A solution of(3S,4S,5S,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yltrifluoromethanesulfonate (100 mg, 0.18 mmol) in methanamine (0.88 mL,1.76 mmol) was stirred at 80° C. overnight. The mixture was purified byflash to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N-methyltetrahydro-2H-pyran-3-amine(60 mg, 76% yield) as a colorless oil. LC-MS (ESI) found: 448 [M+H]⁺.

Step 2: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N-methyltetrahydro-2H-pyran-3-amine(60 mg, 0.13 mmol) in pyridine (1 mL) was added Ac₂O (68 mg, 0.67 mmol).After stirring at rt for 3 h, the mixture was concentrated and theresidue was purified by prep-HPLC (Method B) to giveN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-N-methylacetamide(40 mg, 60.9%) as a colorless oil.LC-MS (ESI) found: 490 [M+H]⁺.

Step 3: A solution ofN-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)-N-methylacetamide(20 mg, 0.04 mmol) in MeOH (2 mL) was added Pd/C (3 mg, 10% wt, 60%wet). The mixture was stirred at rt under a H₂ balloon overnight. Themixture was filtered through a Celite pad, and the filtrate wasconcentrated to giveN-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-N-methylacetamide(4 mg, 44.7%) as a brown oil. LC-MS (ESI) found: 220 [M+H]⁺. ¹H NMR (400MHz, MeOD): δ 4.34 (d, J=1.8 Hz, 1H), 4.14-4.08 (m, 1H), 3.81-3.75 (m,2H), 3.70 (dd, J=4.9, 3.2 Hz, 2H), 3.64-3.58 (m, 1H), 3.40-3.35 (m, 1H),2.80 (s, 3H), 2.21 (s, 3H).

Preparation of(1S,3R,4R,5R,6R)-7,7-dichloro-3-(hydroxymethyl)-2-oxabicyclo[4.1.0]heptane-4,5-diol(Compound A397)

Step 1: To a solution of(2R,3R,4R)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)-3,4-dihydro-2H-pyran(5 g, 12.0 mmol) in CHCl₃ (20 mL) were added TBACI (90 mg, 0.24 mmol)and NaOH aq (8 mL, 50% wt in H₂O). After stirring at 35° C. overnight,the mixture was washed with H₂O, the organic layer was concentrated. Theresidue was purified by flash to give(1S,3R,4R,5R,6R)-4,5-bis(benzyloxy)-3-((benzyloxy)methyl)-7,7-dichloro-2-oxabicyclo[4.1.0]heptane(5 g, 83% yield) as a colorless oil.LC-MS (ESI) found: 499 [M+H]⁺.

Step 2: A solution of(1S,3R,4R,5R,6R)-4,5-bis(benzyloxy)-3-((benzyloxy)methyl)-7,7-dichloro-2-oxabicyclo[4.1.0]heptane(100 mg, 0.44 mmol) in MeOH (2 mL) was added Pd/C (10 mg, 10% wt, 60%wet). The mixture was stirred at rt under a H₂ balloon overnight. Themixture was filtered through a Celite pad, and the filtrate wasconcentrated to give(1S,3R,4R,5R,6R)-7,7-dichloro-3-(hydroxymethyl)-2-oxabicyclo[4.1.0]heptane-4,5-diol(40 mg, 40% yield) as a brown oil. LC-MS (ESI) found: 251 [M+Na]⁺. ¹HNMR (400 MHz, MeOD): δ 3.89 (d, J=9.0 Hz, 1H), 3.82-3.78 (m, 1H), 3.74(t, J=3.1 Hz, 1H), 3.69 (d, J=1.4 Hz, 1H), 3.68 (s, 1H), 3.62 (dd,J=5.1, 1.8 Hz, 1H), 1.82 (dd, J=9.0, 3.3 Hz, 1H).

Preparation of(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-N′—(2,2,2-trifluoroacetyl)tetrahydro-2H-pyran-3-carbohydrazide (Compound A398)

Step 1: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxane-3-carboxylicacid(400 mg, 0.865 mmol) and CDI (0.12 mL, 0.951 mmol) in DCM (20 mL)was added hydrazine hydrate (4.2 mL, 86.5 mmol) at 0° C. The mixture wasstirred at rt for 12 h. the mixture was concentrated and purified bycolumn to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carbohydrazide(140 mg, 34% yield) as yellow solid.

LC-MS (ESI) found: 477 [M+H]⁺.

Step 2: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]oxane-3-carbohydrazide(140 mg, 0.30 mmol) and TFAA (0.08 mL, 0.60 mmol) in MeCN (10 mL) wasadded DIPEA (46 mg, 0.353 mmol) at 0° C. The mixture was stirred at rtfor 2 h. The mixture was concentrated and purified by column to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)-N′—(2,2,2-trifluoroacetyl)tetrahydro-2H-pyran-3-carbohydrazide(135 mg, 80% yield) as yellow oil. LC-MS (ESI) found: 573 [M+H]⁺.

Step 3: To a solution of(4R,5R,6R)-4,5-bis(benzyloxy)-6—[(benzyloxy)methyl]-N′-(trifluoroacetyl)oxane-3-carbohydrazide(10.0 mg, 0.017 mmol) in MeOH (2 mL) was added Pd/C (5 mg, 10% wt, 60%wet). The mixture was stirred at rt for 0.5 h under a H₂ balloon. Themixture was filtered and concentrated to give(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-N′—(2,2,2-trifluoroacetyl)tetrahydro-2H-pyran-3-carbohydrazide(5.1 mg, 95% yield) as white solid. LC-MS (ESI) found: 303 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 4.21 (dd, J=12.2, 0.9 Hz, 1H), 4.00 (dd, J=6.3,3.7 Hz, 1H), 3.79-3.62 (m, 4H), 3.42-3.34 (m, 1H), 2.90 (dd, J=6.2, 2.6Hz, 1H).

Preparation of2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(Compound A399)

Step 1: To a solution of((3S,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl4-methylbenzenesulfonate (400 mg, 0.66 mmol) in DMSO (3 mL) was addedNaCN (98 mg, 1.99 mmol), and the reaction was stirred at roomtemperature for 18 h. The reaction was diluted with EA and water. Theorganic layer was separated, concentrated in vacuo. The residue waspurified with pre-HPLC (Method A) to give2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetonitrile(60 mg, 20% yield).

LC-MS (ESI) found: 458 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 7.48-7.20 (m,15H), 4.81 (d, J=11.1 Hz, 2H), 4.68 (q, J=11.8 Hz, 2H), 4.48 (dt,J=13.7, 11.8 Hz, 3H), 3.99 (dd, J=12.3, 1.8 Hz, 1H), 3.90-3.84 (m, 1H),3.80 (dd, J=4.9, 3.1 Hz, 1H), 3.64-3.52 (m, 4H), 2.91 (qd, J=17.4, 7.2Hz, 2H).

Step 2: To a solution of2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetonitrile(22 mg, 0.048 mmol) in DMSO (4 mL) was added K₂CO₃ (14 mg, 0.096 mmol)and H₂O₂(2 mL). The reaction was stirred at rt for 2 h. The solvent wasremoved, distilled water (10 ml) was added, extracted with EA and thesolvent was removed, purified by flash to give2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(16 mg, 70% yield). LC-MS (ESI) found: 476 [M+H]⁺.

Step 3: To a solution of2-((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)acetamide(20 mg, 0.042 mmol) in MeOH (5 mL) was added Pd/C (3 mg, 10% wt., 60%wet) and HCl (2 mL, 2 M in H₂O). The mixture was stirred at rt 2 h undera H₂ balloon. The mixture was filtered through a Celite pad, and thefiltrate was concentrated to give crude2-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(8 mg, 93% yield). LC-MS (ESI) found: 206[M+H]⁺. ¹H NMR (400 MHz, MeOD):δ 4.62 (dd, J=7.4, 4.5 Hz, 1H), 3.94-3.89 (m, 1H), 3.80-3.73 (m, 2H),3.68-3.61 (m, 2H), 3.52-3.48 (m, 1H), 3.36-3.33 (m, 1H), 2.93 (dd,J=18.3, 6.6 Hz, 1H), 2.83-2.75 (m, 1H).

Preparation of(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethy1681)-N-methyltetrahydro-2H-pyran-3-carboxamide(Compound A400)

Step 1: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (200 mg, 0.432 mmol) in DCM (5 mL) was added SOCl₂ (3 mL, 43 mmol)at rt. The mixture was stirred at 50° C. for 6 h. The mixture wasconcentrated to give crude(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carbonylchloride which was used for next step directly.

Step 2: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carbonylchloride (100 mg, 0.208 mmol) in DCM (2 mL). Methanamine (2.1 mL, 2 M inTHF) was added and the mixture was stirred at rt for 2 h. The mixturewas quenched with H₂O and extracted with DCM, concentrated, purified byprep-HPLC (Method A) to give(4R,5R,6R)-4,5-bis(benzyloxy)-6-(hydroxymethyl)-N-methyloxane-3-carboxamide(20 mg, 25%) as colorless oil. LC-MS (ESI) found: 386 [M+H]⁺.

Step 3: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-(hydroxymethyl)-N-methyltetrahydro-2H-pyran-3-carboxamide(10 mg, 0.026 mmol) in MeOH (2 mL) was added Pd/C (3 mg, 10% wt., 60%wet). The mixture was stirred at rt for 0.5 h under a H₂ balloon. Themixture was filtered and concentrated to give(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)-N-methyltetrahydro-2H-pyran-3-carboxamide(3 mg, 58% yield) as white solid. LC-MS (ESI) found: 206 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 4.08 (dd, J=12.2, 1.0 Hz, 1H), 3.93 (dd, J=6.3, 3.7Hz, 1H), 3.71 (tdd, J=16.0, 11.9, 5.2 Hz, 4H), 3.38-3.34 (m, 1H), 2.79(dd, J=6.2, 2.9 Hz, 1H), 2.75 (s, 3H).

Preparation of methyl(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxylate(Compound A401)

Step 1: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylicacid (A169-1, 100 mg, 0.217 mmol) in MeOH (15 mL) was added H₂SO₄ (19.3mg, 0.108 mmol). The mixture was stirred 60° C. rt for 5 h. The mixturewas concentrated and purified by prep-HPLC (Method A) to give(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylate(55 mg, 53% yield) as yellow oil. LC-MS (ESI) found: 477 [M+H]⁺.

Step 2: To a solution of(3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-carboxylate(15 mg, 0.031 mmol) in MeOH (2 mL) was added Pd/C (4 mg, 10% wt., 60%wet). The mixture was stirred at rt for 0.5 h under a H₂ balloon Themixture was filtered and concentrated to give(3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-carboxylate(6 mg, 92% yield) as yellow oil. LC-MS (ESI) found: 207 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 4.18 (dd, J=12.2, 2.9 Hz, 1H), 3.98 (dd, J=5.6, 3.5Hz, 1H), 3.84-3.75 (m, 2H), 3.74 (s, 3H), 3.71-3.58 (m, 2H), 3.48-3.38(m, 1H), 2.95-2.85 (m, 1H).

Preparation ofN-(2-((((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)amino)ethyl)acetamide(Compound A402)

It was prepared according to the procedure same as that for A209. LC-MS(ESI) found: 263 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 4.05 (d, J=12.4 Hz,1H), 3.96 (dd, J=5.7, 3.3 Hz, 1H), 3.87-3.83 (m, 1H), 3.75 (dd, J=11.5,7.2 Hz, 1H), 3.68-3.62 (m, 2H), 3.59-3.38 (m, 4H), 3.25 (dd, J=12.7, 3.2Hz, 1H), 3.16-3.06 (m, 2H), 2.24-2.15 (m, 1H), 2.00 (s, 3H).

Preparation of1-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)urea(Compound A403)

Step 1: To a solution of((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methanamine(40 mg, 0.089 mmol) in H₂O (10 mL) was added urea (8 mg, 0.13 mmol). Thereaction mixture was stirred at 100° C. overnight.

The resulting mixture was separated and concentrated in vacuo. The crudeproduct was purified by flash chromatography (silica gel, 0˜20% MeOH inDCM) to give1-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)urea(40 mg, 91% yield) as yellow oil. LC-MS (ESI) found: 491 [M+H]⁺.

Step 2: To a solution of1-(((3R,4R,5R,6R)-4,5-bis(benzyloxy)-6-((benzyloxy)methyl)tetrahydro-2H-pyran-3-yl)methyl)urea(40 mg, 0.082 mmol) in MeOH (10 mL) were added Pd/C (10 mg, 10% wt., 60%wet) and HCl (0.1 mL, 1 M in H₂O). The reaction mixture was stirred atrt overnight under a H₂ balloon. The mixture was filtered, the filtratewas concentrated to give1-(((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)methyl)urea(2 mg, 11% yield) as colorless oil. LC-MS (ESI) found: 221 [M+H]⁺. 1HNMR (400 MHz, MeOD): δ 3.96-3.90 (m, 1H), 3.89-3.84 (m, 1H), 3.84-3.77(m, 2H), 3.68 (dd, J=11.6, 4.4 Hz, 1H), 3.58-3.41 (m, 4H), 1.97-1.83 (m,1H).

Allyl Alcohols

Preparation ofN-((2S,3R,4R,5R,6R)-2-(allyloxy)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(Compound A404)

Step 1: To a solution of(2R,3R,4R,5R)-2-amino-3,4,5,6-tetrahydroxyhexanal hydrochloride (20 g,92 mmol) in MeOH (200 mL) was added NaOMe (20.5 ml, 5 M in MeOH) at 0°C., then CF₃COOEt (15 g, 102 mmol) was added. The mixture was stirred atrt for overnight. The mixture was concentrated and washed with i-PrOH togive2,2,2-trifluoro-N-((3R,4R,5R,6R)-2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(20 g, 78% yield) as solid. LC-MS (ESI) found: 298 [M+Na]⁺.

Step 2: To the solution of2,2,2-trifluoro-N-((3R,4R,5R,6R)-2,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)acetamide(20 g, 73 mmol) in pyridine (60 mL) was added Ac₂0 (44 g, 434 mmol) at0° C., then DMAP (1.2 g, 10 mmol) was added at 0° C. The mixture wasstirred at rt overnight. The mixture was concentrated and washed withi-PrOH to give(3R,4R,5R,6R)-6-(acetoxymethyl)-3-(2,2,2-trifluoroacetamido)tetrahydro-2H-pyran-2,4,5-triyltriacetate (20 g, 62% yield) as solid. LC-MS (ESI) found: 442 [M−H]⁻.

Step 3: To the solution of(3R,4R,5R,6R)-6-(acetoxymethyl)-3-(2,2,2-trifluoroacetamido)tetrahydro-2H-pyran-2,4,5-triyltriacetate (5 g, 11.28 mmol) in DCM (50 mL) was added allyl alcohol (2g, 37.76 mmol), then SnCl₄ (7 g, 28.19 mmol) was added at 0° C. Themixture was stirred at rt overnight. the mixture was slowly poured intoice-cold water, the organic layer was separated and washed with NaHCO₃.The organic phase was then dried and concentrated. The residual waspurified by silica gel column to give(2R,3R,4R,5R,6S)-2-(acetoxymethyl)-6-(allyloxy)-5-(2,2,2-trifluoroacetamido)tetrahydro-2H-pyran-3,4-diyldiacetate (2.6 g, 52% yield) as solid. LC-MS (ESI) found: 440 [M−H]⁻.

Step 4: To a solution of(2R,3R,4R,5R,6S)-2-(acetoxymethyl)-6-(allyloxy)-5-(2,2,2-trifluoroacetamido)tetrahydro-2H-pyran-3,4-diyldiacetate (2.6 g, 30.18 mmol) in MeOH (30 mL) was added NaOMe (6 mL, 5 Min MeOH) at 0° C., the mixture was stirred at rt for 3 h. The mixturewas acidified with Amberlite IR120 (H⁺) form and filtered. The filtratewas concentrated to giveN-((2S,3R,4R,5R,6R)-2-(allyloxy)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(1.4 g, 78% yield) as white solid. LC-MS (ESI) found: 316 [M+H]⁺. ¹H NMR(400 MHz, CD₃OD): δ5.98-5.82 (m, 1H), 5.30 (ddd, J=17.3, 3.3, 1.6 Hz,1H), 5.17 (ddd, J=10.5, 2.8, 1.3 Hz, 1H), 4.92 (d, J=3.7 Hz, 1H), 4.29(dd, J=10.9, 3.7 Hz, 1H), 4.24-4.13 (m, 1H), 4.01 (ddt, J=13.2, 6.2, 1.3Hz, 1H), 3.95 (dd, J=10.9, 3.2 Hz, 1H), 3.91 (d, J=2.7 Hz, 1H), 3.85 (t,J=6.1 Hz, 1H), 3.78-3.67 (m, 2H).

Preparation of(2R,3R,4R,5R,6S)-6-(allyloxy)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A405)

Step 1: A solution ofN-((2S,3R,4R,5R,6R)-2-(allyloxy)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)-2,2,2-trifluoroacetamide(A404, 200 mg, 0.6 mmol) in NH₃/MeOH (6 mL, 7 M) was stirred at 80° C.overnight in a sealed tube. The mixture was concentrated and purified byC18 column to give(2R,3R,4R,5R,6S)-6-(allyloxy)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(50 mg, 36% yield) as a white solid. LC-MS (ESI) found: 220 [M+H]⁺.

Step 2: A mixture of(2R,3R,4R,5R,6S)-6-(allyloxy)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(30 mg, 0.1 mmol), 3,5-dichloro-1,2,4-thiadiazole (31 mg, 0.2 mmol) andDIEA (39 mg, 0.3 mmol) in isopropanol (3 mL) was stirred at 80° C.overnight in a sealed tube. The mixture was concentrated and purified bysilica gel column (0-20% methanol in methylene chloride) to give(2R,3R,4R,5R,6S)-6-(allyloxy)-5-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(1.1 mg, 2% yield) as a white solid. LC-MS (ESI) found: 338 [M+H]⁺. ¹HNMR (400 MHz, CD₃OD): δ5.93 (ddd, J=21.9, 10.8, 5.7 Hz, 1H), 5.30 (dd,J=17.2, 1.7 Hz, 1H), 5.16 (dd, J=10.4, 1.6 Hz, 1H), 5.01 (d, J=3.6 Hz,1H), 4.22 (dd, J=13.1, 5.1 Hz, 2H), 4.02 (dd, J=13.0, 6.2 Hz, 1H), 3.92(d, J=2.8 Hz, 1H), 3.89-3.83 (m, 2H), 3.78-3.68 (m, 2H).

Preparation of(2R,3R,4R,5R,6S)-6-(allyloxy)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A406)

Step 1: To a mixture of(2R,3R,4R,5R,6S)-6-(allyloxy)-5-amino-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(30 mg, 0.14 mmol) in i-PrOH (2 mL) was added2-chloro-4-(trifluoromethyl)pyrimidine (0.03 mL, 0.27 mmol) and DIEA(0.06 mL, 0.41 mmol) at rt under N₂. After stirring at 120° C.overnight, the reaction mixture was concentrated and purified byprep-TLC to afford(2R,3R,4R,5R,6S)-6-(allyloxy)-2-(hydroxymethyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(21 mg, 41% yield) as a white solid. LC-MS (ESI) found: 366 [M+H]⁺. ¹HNMR (400 MHz, CD₃OD): δ8.52 (d, J=4.7 Hz, 1H), 6.91 (d, J=4.9 Hz, 1H),5.88 (s, 1H), 5.27 (d, J=16.9 Hz, 1H), 5.10 (d, J=10.6 Hz, 1H), 5.02 (s,1H), 4.53 (s, 1H), 4.21 (dd, J=13.1, 5.0 Hz, 1H), 4.01-3.86 (m, 4H),3.79-3.70 (m, 2H).

Preparation of(2R,3R,4R,5R)-5-(aminomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(Compound A425)

Step 1: To a solution of(2R,3R,4R,5R)-5-(azidomethyl)-3,4-bis(benzyloxy)-2-((benzyloxy)methyl)tetrahydro-2H-pyran(50 mg, 0.11 mmol) in MeOH (5 mL) were added Pd/C (5 mg, 10% wt, 60%wet) and HCl (0.1 mL, 1 N in H₂O). The reaction mixture was stirred atrt overnight under a balloon of H₂. The mixture was filtered andconcentrated in vacuo to give(2R,3R,4R,5R)-5-(aminomethyl)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(16 mg, 86% yield) as yellow oil. LC-MS (ESI) found: 178 [M+H]⁺. ¹H NMR(400 MHz, CD3OD): δ4.09-4.02 (m, 1H), 3.96 (dd, J=5.7, 3.3 Hz, 1H), 3.85(d, J=2.7 Hz, 1H), 3.76-3.70 (m, 1H), 3.69-3.61 (m, 2H), 3.43-3.33 (m,2H), 3.26 (dd, J=13.0, 3.7 Hz, 1H), 2.22-2.05 (m, 1H).

The following compounds below were prepared according to the proceduresame as A287:

ID Characterization data Starting Material

Yield: 2.6 mg, 3%, white solid. LC-MS (ESI) found: 325 [M + H]⁺. ¹H NMR(400 MHz, CD₃OD): δ 6.00 (s, 1H), 5.29 (d, J = 0.9 Hz, 1H), 4.25 (dd, J= 17.2, 7.1 Hz, 1H), 3.93 (d, J = 11.4 Hz, 1H), 3.90 (d, J = 4.3 Hz,1H), 3.89 (s, 3H), 3.83 (d, J = 11.4 Hz, 1H), 3.77 (dd, J = 8.8, 4.5 Hz,2H), 3.70 (d, J = 7.9 Hz, 1H).

Yield: 1.4 mg, 2%, white solid. LC-MS (ESI) found: 338 [M + H]⁺. ¹H NMR(400 MHz, CD₃OD): δ 8.16 (s, 1H), 8.01 (s, 1H), 5.35 (d, J = 1.4 Hz,1H), 4.21 (dd, J = 9.9, 1.2 Hz, 1H), 3.94 (d, J = 10.2 Hz, 1H), 3.92 (d,J = 3.0 Hz, 1H), 3.84 (dd, J = 9.6, 4.5 Hz, 2H), 3.81 (d, J = 9.0 Hz,1H), 3.72 (d, J = 7.9 Hz, 1H).

Linkers:

Preparation ofN-((1S,2R,3R,4R,5S)-1-(13-azido-2,5,8,11-tetraoxatridecyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)-2,2,2-trifluoroacetamide(Compound A407)

Step 1: The solution ofN-((1S,2R,3R,4R,5S)-2,3-dihydroxy-1-(hydroxymethyl)-6,8-dioxabicyclo[3.2.1]octan-4-yl)-2,2,2-trifluoroacetamidein DMF (20 mL) was added CSA (0.81 g, 3.48 mmol) and2,2-dimethoxypropane (7.54 g, 72.43 mmol). The reaction mixture wasstirred at 80° C. overnight and quenched with TEA. Then the solvent wasremoved under vacuum. The residue was purified by flash chromatography(DCM:MeOH=10:1) to give 2,2,2-trifluoro-N-((3 aR,4 S,7 S, 8R,8aR)-4-(hydroxymethyl)-2,2-dimethylhexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-8-yl)acetamide (2 g, 68% yield) as a colorless oil. LC-MS(ESI) found: 328 [M+H]⁺.

Step 2: To a suspension of2,2,2-trifluoro-N-((3aR,4S,7S,8R,8aR)-4-(hydroxymethyl)-2,2-dimethylhexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-8-yl)acetamide(2.27 g, 6.94 mmol) in dry DMF (30 mL) was added NaH (0.33 g, 8.32 mmol,60% weight in mineral oil) was added. After stirring at rt for 1.5 h,2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate(2.85 g, 7.63 mmol) was added. The mixture was stirred at 60° C. for 16h, NH₄Cl (aq) was added, the solvent was removed, and purified byprep-HPLC (Method B) to giveN-((3aR,4S,7S,8R,8aR)-4-(13-azido-2,5,8,11-tetraoxatridecyl)-2,2-dimethylhexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-8-yl)-2,2,2-trifluoroacetamide(650 mg, 18% yield). LC-MS (ESI) found: 529 [M+H]⁺.

Step 3: A solution ofN-((3aR,4S,7S,8R,8aR)-4-(13-azido-2,5,8,11-tetraoxatridecyl)-2,2-dimethylhexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-8-yl)-2,2,2-trifluoroacetamide(30 mg, 0.057 mmol) in AcOH (3.2 mL), MeOH (1 mL) and H₂O (1 mL) wasstirred at 70° C. for 14 h. Solvent was evaporated and the residue wasco-evaporated twice with toluene. The residue was purified by prep-HPLC(Method B) to giveN-((1S,2R,3R,4R,5S)-1-(13-azido-2,5,8,11-tetraoxatridecyl)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-4-yl)-2,2,2-trifluoroacetamide(10 mg, 39% yield). LC-MS (ESI) found: 489 [M+H]⁺. ¹H NMR (400 MHz,CDCl₃): δ 6.51 (d, J=8.5 Hz, 1H), 5.38 (d, J=1.3 Hz, 1H), 4.81 (d, J=4.7Hz, 1H), 4.18-4.05 (m, 3H), 3.81 (d, J=8.2 Hz, 1H), 3.76-3.63 (m, 16H),3.58 (s, 1H), 3.42 (td, J=4.7, 1.9 Hz, 2H), 3.18 (d, J=10.1 Hz, 1H)

Preparation of13,13-bis((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (Compound A408)

Step 1: To a solution of2-[2-(2-{2—[(2,4-dinitrophenyl)amino]ethoxy}ethoxy)ethoxy]acetic acid (3g, 8.04 mmol) in DMF (30 mL) were added HATU (3.97 g, 10.45 mmol) andDIEA (3.98 mL, 24.11 mmol). The reaction mixture was stirred at icewater for 0.5 hours, then di-tert-butyl3,3′-((2-amino-2-((3-(tert-butoxy)-3-oxopropoxy)methyl)propane-1,3-diyl)bis(oxy))dipropionate(4.47 g, 8.84 mmol) was added. The reaction mixture was stirred at rtfor 16 h and purified by prep-HPLC (Method B) to give tert-butyl13,13-bis((3-(tert-butoxy)-3-oxopropoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oate(4 g, 89%). LC-MS (ESI) found: 861 [M+H]⁺.

Step 2: To a solution of tert-butyl13,13-bis((3-(tert-butoxy)-3-oxopropoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oate(500 mg, 0.58 mmol) in DCM (8 mL) was added TFA (0.8 mL).The reactionwas stirred at rt for 18 h. The reaction was concentrated in vacuo andthe residue was purified by flash (C18) (water/CH₃CN) to give13,13-bis((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (110 mg, 75%). LC-MS (ESI) found: 693 [M+H]⁺. ¹H NMR (400 MHz,MeOD): δ 9.04 (d, J=2.7 Hz, 1H), 8.29 (dd, J=9.6, 2.7 Hz, 1H), 7.23 (d,J=9.6 Hz, 1H), 3.88 (s, 2H), 3.82 (t, J=5.3 Hz, 2H), 3.74-3.64 (m, 22H),2.51 (t, J=6.1 Hz, 6H).

Preparation of(2S,3R,4R,5R)-N-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-carboxamide(Compound A409)

Step 1: To a solution of 1-chloro-2,4-dinitrobenzene (1.0 g, 4.94 mmol)in H₂O (20 mL) was added NaHCO₃ (0.4 g, 5.13 mmol) and tert-butyl(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)carbamate (0.5 g, 1.71 mmol)at 0° C. in portions. The mixture was stirred at 95° C. for 2 h. Themixture was adjust pH to 6 and extracted with DCM, washed with brine,concentrated and purified by silica column to give(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl)carbamate(710 mg, 90% yield) as a yellow oil. LC-MS (ESI) found: 459 [M+H]⁺.

Step 2: To a solution of tert-butyl(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl)carbamate(100 mg, 0.22 mmol) in DCM (5 mL) was added TFA (0.8 mL, 10.9 mmol) andstirred at rt for 2 h. The mixture was concentrated to give crudeN-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-2,4-dinitroaniline (75mg, 96% yield) as a yellow oil. LC-MS (ESI) found: 359 [M+H]⁺.

Step 3: To a solution ofN-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-2,4-dinitroaniline and(2S,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-carboxylicacid in DCM is added EDCI, HOBt, NMM at 0° C. The mixture is stirred atrt for 2 h. The mixture is purified by prep-HPLC (Method A) to give(2S,3R,4R,5R)-N-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-carboxamide.

Preparation of6-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethoxy)benzo[d]thiazole-2-carbonitrile(Compound A410)

Step 1: To a solution of(2R,3R,4R,5R)-2-(aminomethyl)-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-3,4-diolin DMF is added2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl4-methylbenzenesulfonate and Cs₂CO₃. The reaction mixture is stirred atrt overnight. The resulting mixture is extracted with EA, washed withH₂O and brine, and dried over Na₂SO₄. The organic layer is separated andconcentrated in vacuo. The crude product is purified by flashchromatography (silica gel, 0-50% EA in PE) to give(2R,3R,4R,5R)-2-(13-((2,4-dinitrophenyl)amino)-5,8,11-trioxa-2-azatridecyl)-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-3,4-diol.

Preparation ofN-(1-((1S,2R,3R,4R,5S)-2,3-dihydroxy-4-(2,2,2-trifluoroacetamido)-6,8-dioxabicyclo[3.2.1]octan-1-yl)-2,5,8,11-tetraoxatridecan-13-yl)but-3-ynamide(Compound A411)

Preparation of(2R,3R,4R,5S)-5-((2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)amino)-2-(13-((2,4-dinitrophenyl)amino)-5,8,11-trioxa-2-azatridecyl)tetrahydro-2H-pyran-3,4-diol(Compound A412)

Step 1: To a solution of 2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethan-1-ol(1.2 g, 6.21 mmol) in H₂O (20 mL) was added NaHCO₃ (1.6 g, 18.63 mmol)and 1-chloro-2,4-dinitrobenzene (2.5 g, 12.42 mmol) at 0° C. The mixturewas stirred at 95° C. for 2 h. The mixture was adjust PH to 6 andextracted with DCM, washed with brine, concentrated and purified bysilica gel column to give2-2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethan-1-ol(2.1 g, 941%) as a yellow oil. LC-MS (ESI) found: 360 [M+H]⁺.

Step 2: To a solution of2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethan-1-ol(1.0 g, 2.78 mmol) in DCM (20 mL) was added TsCl (635 mg, 3.340 mmol)and TEA (1.16 mL, 8.35 mmol) at 0° C. in portions. The mixture wasstirred at rt for 2 h. The mixture was concentrated and purified bysilica gel column to give2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl4-methylbenzenesulfonate (1.2 g, 87%) as a yellow oil. LC-MS (ESI)found: 514 [M+H]⁺.

Step 3: To a solution of2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl4-methylbenzenesulfonate in DMF is added(2R,3R,4R,5S)-2-(aminomethyl)-5-((2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)amino)tetrahydro-2H-pyran-3,4-dioland K₂CO₃ at 0° C. The mixture is stirred at 100° C. for 12 h. Themixture is concentrated and purified by silica gel column to give(2R,3R,4R,5S)-5-((2-chloropyrrolo[2,1-f][1,2,4]triazin-4-yl)amino)-2-(13-((2,4-dinitrophenyl)amino)-5,8,11-trioxa-2-azatridecyl)tetrahydro-2H-pyran-3,4-diol.

Preparation of(2R,3R,4R,5S)-2-(13-azido-2,5,8,11-tetraoxatridecyl)-5-(methyl(4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound A413)

Preparation of3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(((2R,3R,4R,5S)-5-((3-amino-1,2,4-thiadiazol-5-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methyl)propanamide)(Compound A414)

Preparation of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(Compound A415)

Step 1: A solution of 6-azidohexanoic acid (0.54 g, 3.45 mmol) and HATU(1.3 g, 3.45 mmol) in DMF (10 mL) was stirred at rt for 30 min.Di-tert-butyl 3,3′-((2-aminopropane-1,3-diyl)bis(oxy))dipropionate (1 g,2.88 mmol) and DIPEA (0.95 mL, 5.76 mmol) were added at rt. The reactionwas stirred overnight. The resulting mixture was diluted with DCM (100mL),washed with H₂O (50 mL×2) and brine (20 mL),dried over Na₂SO₄. Theorganic layer was separated and concentrated in vacuo. The crude productwas purified by flash chromatography (silica gel, 0˜80% EA in PE) togive di-tert-butyl3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))dipropionate (800mg, 93% yield) as colorless oil. LC-MS (ESI) found: 487 [M+H]⁺. ¹H NMR(400 MHz, CDCl₃): δ 6.28 (d, J=8.2 Hz, 1H), 4.19-4.11 (m, 1H), 3.74-3.62(m, 4H), 3.58 (dd, J=9.6, 4.0 Hz, 2H), 3.40 (dd, J=9.6, 6.1 Hz, 2H),3.26 (t, J=6.9 Hz, 2H), 2.52-2.38 (m, 4H), 2.20 (t, J=7.5 Hz, 2H),1.67-1.63 (m, 4H), 1.47-1.37 (m, 20H).

Step 2: A solution of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))dipropionate (800mg, 1.64 mmol) in DCM (6 mL) was added TFA (2 mL, 26.93 mmol) dropwiseat 0° C. The reaction was stirred at rt for 3 h. The resulting mixturewas filtered and concentrated in vacuo.

The crude3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))dipropionic acid(470 mg, 76% yield) was used to next step with no further purification.LC-MS (ESI) found: 375 [M+H]⁺. ¹H NMR (400 MHz, DMSO-d₆): δ 12.14 (s,2H), 7.64 (d, J=8.2 Hz, 1H), 4.00-3.86 (m, 1H), 3.57 (t, J=6.3 Hz, 4H),3.34 (d, J=5.8 Hz, 4H), 3.29 (d, J=6.9 Hz, 2H), 2.43 (t, J=6.3 Hz, 4H),2.07 (t, J=7.3 Hz, 2H), 1.56-1.43 (m, 4H), 1.33-1.23 (m, 2H).

Step 3: A solution of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))dipropionic acid(70 mg, 0.19 mmol) and HATU (163 mg, 0.43 mmol) in DMF (5 mL) wasstirred at rt for 30 min.(2R,3R,4R,5S)-2-(13-amino-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(208 mg, 0.43 mmol) and DIPEA (73 mg, 0.56 mmol) were added at rt. Thereaction was stirred overnight. The resulting mixture was concentratedin vacuo. The crude product was purified by flash chromatography (silicagel, 0˜10% MeOH in DCM) to give3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(65 mg, 28% yield) as white solid. LC-MS (ESI) found: 654 [M+2H]²⁺. ¹HNMR (400 MHz, MeOD): δ 8.51 (d, J=4.8 Hz, 2H), 6.90 (d, J=4.9 Hz, 2H),4.36 (td, J=10.6, 5.3 Hz, 2H), 4.16-4.04 (m, 3H), 3.92 (d, J=2.9 Hz,2H), 3.73-3.60 (m, 36H), 3.56 (dt, J=10.6, 3.0 Hz, 6H), 3.51-3.43 (m,4H), 3.38 (t, J=5.5 Hz, 4H), 3.17 (t, J=10.9 Hz, 2H), 2.45 (dd, J=6.7,5.4 Hz, 4H), 2.23 (t, J=7.5 Hz, 2H), 1.67-1.57 (m, 4H), 1.44-1.36 (m,2H). ¹⁹F NMR (377 MHz, MeOD): δ-72.30 (s).

Preparation of2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetic acid(Compound A416)

Step 1: To a solution of 2-(2-(2-aminoethoxy)ethoxy)ethan-1-ol (5.0 g,33 mmol) in EtOH (60 mL) was added TEA (61.5 mL, 442 mmol) anddi-tert-butyl dicarbonate (8.6 mL, 40 mmol) at 0° C. in portions. Themixture was vigorously stirred and allowed to warm up to roomtemperature slowly overnight (16 h). The solvents were evaporated undervacuum and the residue was purified by column chromatography (SiO₂,solvent gradient: DCM to 1:9 MeOH/DCM) to give tert-butyl(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)carbamate as a colorless oil (4.5 g,55% yield). LC-MS (ESI) found: 250 [M+H]⁺.

Step 2: To a solution of tert-butyl(2-(2-(2-hydroxyethoxy)ethoxy)ethyl)carbamate as a colorless oil (72 g,288 mmol) in THE (100 mL) was added 2-iodoacetic acid (160 g, 866 mmol)and NaOH (69 g, 1.7 mol) at 0° C. in portions. The mixture was stirredat rt for 2 days. The solvent was removed under vacuum, and then asolution of NaOH in water was added. DCM was used to wash the mixture.The aqueous phase was then acidified with 3 N HCl solution with vigorousstirring until pH 4. Extraction with DCM and concentration give2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecan-16-oic acid as ayellowish oil (80 g, 90% yield). LC-MS (ESI) found: 308 [M+H]⁺.

Step 3: To a solution of2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadecan-16-oic acid as ayellowish oil (42 g, 136 mmol) in DCM (250 mL) was added TFA (51 mL, 683mmol) at 0° C. The mixture was stirred at rt for 2 h. The mixture wasconcentrated to give crude 2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)aceticacid (28 g, 99% yield) as a white solid. LC-MS (ESI) found: 208 [M+H]⁺.

Step 4: To a solution of 2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)aceticacid (28 g, 135 mmol) in H₂O (300 mL) was added NaHCO₃ (34 g, 405 mmol)and 1-chloro-2,4-dinitrobenzene (41 g, 203 mmol) at 0° C. in portions.The mixture was stirred at 90° C. overnight. The mixture was adjust pHto 6, extracted with DCM, washed with brine, concentrated and purifiedby silica gel column to give2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetic acid(20 g, 40% yield) as a yellow oil. LC-MS (ESI) found: 374 [M+H]⁺. ¹H NMR(400 MHz, MeOD): δ 9.03 (d, J=2.7 Hz, 1H), 8.28 (dd, J=9.6, 2.7 Hz, 1H),7.22 (d, J=9.6 Hz, 1H), 4.10 (s, 2H), 3.81 (t, J=5.3 Hz, 2H), 3.75-3.61(m, 1OH).

Preparation of6-azido-N-(1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)hexanamide(Compound A417)

Step 1: To a solution of benzyl(1,3-bis(prop-2-yn-1-yloxy)propan-2-yl)carbamate (80 mg, 0.27 mmol) and(2R,3R,4R,5S)-2-(13-azido-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(A419, 300 mg, 0.58 mmol) in MeOH (3 mL) were added CuSO₄ (4.2 mg, 0.03mmol) and THPTA (20 mg, 0.004 mmol) in H₂O (0.5 mL) and Na ascorbate (11mg, 0.05 mmol) in H₂O (0.5 mL). The mixture was stirred at rt overnightand concentrated. The residue was purified by flash to give benzyl(1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-(1,3-yl)-¹H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)carbamate(A420, 320 mg, 91% yield) as colorless oil. LC-MS (ESI) found: 1322[M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 8.49 (d, J=4.8 Hz, 2H), 8.01 (s, 2H),7.47-7.19 (m, 5H), 6.88 (d, J=4.9 Hz, 2H), 5.07 (s, 2H), 4.57 (dd,J=9.5, 4.6 Hz, 8H), 4.35 (d, J=5.1 Hz, 2H), 4.06 (dd, J=10.9, 5.2 Hz,2H), 3.95-3.85 (m, 7H), 3.69-3.63 (m, 6H), 3.62-3.51 (m, 30H), 3.13 (t,J=10.9 Hz, 2H).

Step 2: A solution of benzyl(1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)carbamate(300 mg, 0.23 mmol) and Pd/C (30 mg, 10% wt, 60% wet) in MeOH (5 mL) wasstirred at rt under a H₂ balloon overnight. The mixture was filteredthrough a Celite pad, and the filtrate was concentrated to give(2R,2′R,3R,3′R,4R,4′R,5S,5'S)-2,2′-(((((2-aminopropane-1,3-diyl)bis(oxy))bis(methylene))bis(1H-1,2,3-triazole-4,1-diyl))bis(2,5,8,11-tetraoxatridecane-13,1-diyl))bis(5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol)(160 mg, 60% yield) as colorless oil. LC-MS (ESI) found: 1188 [M+H]⁺.

Step 3: To a solution of 6-azidohexanoic acid (13 mg, 0.08 mmol) in DMF(3 mL) was added HATU (35 mg, 0.09 mmol) and DIPEA (22 mg, 0.17 mmol).After stirring at rt for 30 min,(2R,2′R,3R,3′R,4R,4′R,5S,5'S)-2,2′-(((((2-aminopropane-1,3-diyl)bis(oxy))bis(methylene))bis(1H-1,2,3-triazole-4,1-diyl))bis(2,5,8,11-tetraoxatridecane-13,1-diyl))bis(5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol)(147 mg, 0.12 mmol) was added. The mixture was stirred at rt under N₂overnight. The mixture was concentrated and the residue was purified byprep-HPLC (Method A) to give6-azido-N-(1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)hexanamide(36 mg, 33% yield) as white solid. LC-MS (ESI) found: 1327 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.50 (d, J=4.8 Hz, 2H), 8.02 (s, 2H), 6.89 (d,J=4.9 Hz, 2H), 4.67-4.53 (m, 8H), 4.35 (td, J=10.4, 5.2 Hz, 2H), 4.19(p, J=5.5 Hz, 1H), 4.07 (dd, J=10.9, 5.2 Hz, 2H), 3.89 (t, J=4.9 Hz,5H), 3.69-3.50 (m, 37H), 3.26 (t, J=6.8 Hz, 2H), 3.14 (t, J=10.9 Hz,2H), 2.21 (t, J=7.4 Hz, 2H), 1.66-1.52 (m, 4H), 1.45-1.27 (m, 2H).

Preparation of3,3′-((2-(2-(2-(2-(2-(4-((2-phenylpiperidin-1-yl)methyl)-1H-indol-1-yl)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((1S,2R,3R,4R,5S)-2,3-dihydroxy-4-(2,2,2-trifluoroacetamido)-6,8-dioxabicyclo[3.2.1]octan-1-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide(Compound A421)

Preparation of3,3′-((2-aminopropane-1,3-diyl)bis(oxy))bis(N-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(Compound A422)

Step 1: To a solution of3,3′-((2-(((benzyloxy)carbonyl)amino)propane-1,3-diyl)bis(oxy))dipropionicacid (0.72 g, 1.96 mmol) and HATU (2.23 g, 5.88 mmol) in DMF (20 mL) wasadded DIPEA (1.0 g, 7.84 mmol) and(2R,3R,4R,5S)-2-(13-amino-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(2.1g, 4.33 mmol) at rt. The reaction mixture was stirred at rt forovernight. The resulting mixture was separated and concentrated invacuo. The crude product was purified by flash chromatography (C18,0˜80%, MeOH in H₂O) to give benzyl(1,39-bis((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-15,25-dioxo-2,5,8,11,18,22,29,32,35,38-decaoxa-14,26-diazanonatriacontan-20-yl)carbamate(A423, 1.2 g, 48% yield) as white solid. LC-MS (ESI) found: 1303[M+H]⁺.¹H NMR (400 MHz, MeOD): δ 8.50 (d, J=4.8 Hz, 2H), 7.41-7.25 (m, 5H),6.89 (d, J=4.9 Hz, 2H), 5.07 (s, 2H), 4.43-4.27 (m, 2H), 4.08 (dd,J=10.9, 5.2 Hz, 2H), 3.91 (d, J=3.0 Hz, 2H), 3.88-3.82 (m, 1H),3.72-3.55 (m, 36H), 3.55-3.42 (m, 8H), 3.35 (t, J=5.5 Hz, 4H), 3.15 (t,J=10.9 Hz, 2H), 2.43 (t, J=6.1, 4H).

Step 2: To a solution of benzyl(1,39-bis((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-15,25-dioxo-2,5,8,11,18,22,29,32,35,38-decaoxa-14,26-diazanonatriacontan-20-yl)carbamate(1.2g, 0.92 mmol) in MeOH (10 mL) was added Pd/C (120 mg, 10% wt, 60%wet) at rt. The reaction mixture was stirred at rt under a H₂ balloonovernight. The resulting mixture was separated and concentrated invacuo. The crude product was purified by flash chromatography (C18,0˜40%, MeOH in H₂O) to give3,3′-((2-aminopropane-1,3-diyl)bis(oxy))bis(N-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(800 mg, 74% yield) as white solid. LC-MS (ESI) found: 1169[M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 8.51 (d, J=4.7 Hz, 2H), 6.89 (d, J=4.9 Hz, 2H),4.36 (d, J=5.0 Hz, 2H), 4.08 (dd, J=11.0, 5.1 Hz, 2H), 3.89 (t, J=17.0Hz, 2H), 3.79-3.52 (m, 40H), 3.46 (dt, J=20.7, 10.3 Hz, 2H), 3.37 (td,J=8.5, 4.9 Hz, 6H), 3.13 (dt, J=10.3, 8.0 Hz, 3H), 2.46 (t, J=6.0 Hz,4H).

Preparation of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((1S,2R,3R,4R,5S)-4-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-1-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(Compound A424)

Step 1: A solution of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))dipropionic acid(28 mg, 0.08 mmol) and HATU (62.7 mg, 0.17 mmol) in DMF (3 mL) wasstirred at rt for 30 min.N-((3aR,4S,7S,8R,8aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-2,2-dimethylhexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-8-yl)-3-chloro-1,2,4-thiadiazol-5-amine(98 mg, 0.19 mmol) and DIPEA (0.05 mL, 0.3 mmol) were added at rt. Thereaction was stirred overnight. The resulting mixture was concentratedin vacuo. The crude product was purified by flash chromatography (silicagel, 0˜10% MeOH in DCM) to give3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((3aR,4S,7S,8R,8aR)-8-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2,2-dimethyltetrahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4(5H)-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(50 mg, 48% yield) as a colorless oil. LC-MS (ESI) found: 1389 [M+H]⁺.

Step 2: To a solution of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((3aR,4S,7S,8R,8aR)-8-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2,2-dimethyltetrahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4(5H)-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(50 mg, 0.04 mmol) in THE (3 mL) was added HCl (1 mL, 2 M in H₂O). Thereaction was stirred at rt overnight. The crude product was purified byflash chromatography (silica gel, 0-10% MeOH in DCM) to give3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((1S,2R,3R,4R,5S)-4-((3-chloro-1,2,4-thiadiazol-5-yl)amino)-2,3-dihydroxy-6,8-dioxabicyclo[3.2.1]octan-1-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(30 mg, 64% yield) as a white solid. LC-MS (ESI) found: 1309 [M+H]⁺. ¹HNMR (400 MHz, MeOD): δ 5.39 (s, 2H), 4.14-4.09 (m, 1H), 4.00 (d, J=9.6Hz, 2H), 3.93 (d, J=4.1 Hz, 2H), 3.83 (d, J=8.0 Hz, 2H), 3.77 (dd,J=9.6, 4.2 Hz, 2H), 3.73-3.60 (m, 35H), 3.56 (t, J=5.5 Hz, 4H),3.52-3.45 (m, 4H), 3.39 (t, J=5.5 Hz, 4H), 3.28 (s, 1H), 2.46 (dd,J=6.6, 5.4 Hz, 4H), 2.23 (t, J=7.4 Hz, 2H), 1.67-1.57 (m, 4H), 1.44-1.36(m, 2H).

Degraders

Preparation of Compound 28((S)-2-((S)-33-((1H-imidazol-5-yl)methyl)-1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-20,20-bis(1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22,31-trioxo-2,5,8,11,18,25,28-heptaoxa-14,21,32-triazatetratriacontan-34-amido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide)

Step 1: To a solution of ethane-1,2-diol (1.8 mL, 32.2 mmol) andtert-butyl prop-2-enoate (11.7 5 mL, 80.6 mmol) in DCM (65 mL) and H₂O(2.6 mL) was added Bu₄NBr (6.2 g, 19.3 mmol) and sodium hydroxide (2.6g, 65.0 mmol) at room temperature. The mixture was stirred at roomtemperature for 12 h. Then the mixture was extracted by DCM and theorganic phase was concentrated. The residual was then purified by flashchromatography (silica gel, 0˜15% PE in EA) to give di-tert-butyl3,3′-(ethane-1,2-diylbis(oxy))dipropionate (4.3 g, 42% yield). LC-MS 10(ESI) found: 319 [M+H]⁺.

Step 2: To a mixture of di-tert-butyl3,3′-(ethane-1,2-diylbis(oxy))dipropionate (4.3 g, 13.51 mmol) in DCM(28 mL) was added TFA (5.6 mL). The mixture was stirred for 4 h at roomtemperature. The mixture was concentrated to give crude3,3′-(ethane-1,2-diylbis(oxy))dipropionic acid (2.8 g, 94% yield). LC-MS(ESI) found: 207 [M+H]⁺.

Step 3: To a solution of 3,3′-(ethane-1,2-diylbis(oxy))dipropionic acid(2.8 g, 13.6 mmol) and (4- phenylphenyl)methanol (2.50 g, 13.58 mmol)were added PTSA (0.02 g, 0.14 mmol) under N₂. The mixture was stirred at140° C. for 3 h. Then NaHCO₃ was added, and the mixture was extracted byDCM and the organic phase was concentrated. The residual was thenpurified by flash chromatography (silica gel, 0˜50% methanol indichloromethane) to give3-(2-(3-([1,1′-biphenyl]-4-ylmethoxy)-3-oxopropoxy)ethoxy)propanoic acid(0.88 g, 17% yield). LC-MS (ESI) found: 373 [M+H]⁺.

Step 4: To a solution of3-(2-(3-([1,1′-biphenyl]-4-ylmethoxy)-3-oxopropoxy)ethoxy)propanoic acid(500 mg, 1.34 mmol) and di-tert-butyl3,3′-((2-amino-2-((3-(tert-butoxy)-3-oxopropoxy)methyl)propane-1,3-diyl)bis(oxy))dipropionate(792 mg, 1.61 mmol) in DMF (5 mL) was added HATU (765 mg, 2.01 mmol) andDIPEA (0.7 mL, 4.02 mmol) at room temperature. The mixture was stirredat room temperature for 12 h. Then the mixture was extracted by DCM andthe organic phase was concentrated. The residual was then purified byflash chromatography (silica gel, 0˜48% ethyl acetate in petroleumether) to give 17-([1,1′-biphenyl]-4-ylmethyl) 1-(tert-butyl)6,6-bis((3-(tert-butoxy)-3-oxopropoxy)methyl)-8-oxo-4,11,14-trioxa-7-azaheptadecanedioate(1 g, 87% yield) as a solid. LC-MS (ESI) found: 860 [M+H]⁺.

Step 5: To a mixture of 17-([1,1′-biphenyl]-4-ylmethyl) 1-(tert-butyl)6,6-bis((3-(tert-butoxy)-3-oxopropoxy)methyl)-8-oxo-4,11,14-trioxa-7-azaheptadecanedioate(1 g, 1.16 mmol) in DCM (10 mL) was added TFA (2 mL). The mixture wasstirred for 4 h at room temperature. The mixture was concentrated togive crude1-([1,1′-biphenyl]-4-yl)-14,14-bis((2-carboxyethoxy)methyl)-3,12-dioxo-2,6,9,16-tetraoxa-13-azanonadecan-19-oicacid (620 mg, 77%). LC-MS (ESI) found: 692 [M+H]⁺.

Step 6: To a solution of1-([1,1′-biphenyl]-4-yl)-14,14-bis((2-carboxyethoxy)methyl)-3,12-dioxo-2,6,9,16-tetraoxa-13-azanonadecan-19-oicacid (40 mg, 0.060 mmol) in DMF (5 mL) was added HATU (102 mg, 0.27mmol). After stirring at rt for 30 min,N-((3aR,4R,6R,7R,7aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(103 mg, 0.23 mmol) and DIPEA (77 mg, 0.60 mmol) were added. The mixturewas stirred at rt for 8 h, then diluted with ethyl acetate and washedwith water. The organic layer was dried over sodium sulfate, filtered,concentrated under reduced pressure to give a crude product, which waspurified by column to give [1,1′-biphenyl]-4-ylmethyl1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,25,28-heptaoxa-14,21-diazahentriacontan-31-oate(60 mg, 50% yield). LC-MS (ESI) found: 1989 [M+H]⁺.

Step 7: To a solution of [1,1′-biphenyl]-4-ylmethyl1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,25,28-heptaoxa-14,21-diazahentriacontan-31-oate(60 mg, 0.030 mmol) in MeOH (5 mL), Pd/C (10 mg, 10% wt, 60% wet) wasadded. The reaction mixture was stirred at rt overnight under a H₂balloon. The mixture was filtered and concentrated to give1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,25,28-heptaoxa-14,21-diazahentriacontan-31-oicacid (14 mg, 26% yield) as yellow oil. LC-MS (ESI) found: 1822 [M+H]⁺.

Step 8: To a solution of1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,25,28-heptaoxa-14,21-diazahentriacontan-31-oicacid (14 mg, 0.008 mmol) in DMF (1 mL) was added HATU (3.8 mg, 0.010mmol). After stirring at rt for 30 min,(S)-2-((S)-2-amino-3-(1H-imidazol-5-yl)propanamido)-N1-((S)-1-(((3S,6R,11R,14S,175,205,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(15 mg, 0.008 mmol) and DIPEA (2 mg, 0.015 mmol) were added. The mixturewas stirred at rt for 8 h, then the mixture was purified by pre-HPLC(Method B) to give 28-11 (20 mg, 71% yield). LC-MS (ESI) found: 1219[M+3H]³⁺.

Step 9: To a solution of 28-11 (20 mg, 0.014 mmol) in THF (0.5 mL) wasadded HCl (0.1 mL, 2 N in H₂O). The reaction was stirred at rt for 16 h.The reaction was concentrated in vacuo. The residue was purified withprep-HPLC (Method B) to give(S)-2-((S)-33-((1H-imidazol-5-yl)methyl)-1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-20,20-bis(1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22,31-trioxo-2,5,8,11,18,25,28-heptaoxa-14,21,32-triazatetratriacontan-34-amido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,10H-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(Compound 28, 8.2 mg, 40% yield). LC-MS (ESI) found: 1179 [M+3H]3.

Preparation of Compound 29:N-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-3-((14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide

Step 1: To a solution of2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetic acid(200 mg, 0.54 mmol) in DCM (5 mL) were added HATU (306 mg, 0.81 mmol)and DIPEA (207 mg, 1.61 mmol) at 0° C. in portions. After stirring at 0°C. for 30 min, tert-butyl 3-(2-aminoethoxy)propanoate (152 mg, 0.81mmol) was added. The mixture was stirred at rt for 12 h. The mixture wasextracted with DCM and washed with brine and concentrated and purifiedby silica gel column to give tert-butyl1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oate(280 mg, 96% yield) as yellow oil. LC-MS (ESI) found: 545 [M+H]⁺.

Step 2: To a solution of tert-butyl1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oate(100 mg, 0.18 mmol) in DCM (3 mL) was added TFA (0.5 mL) at 0° C. Themixture was stirred at rt for 12 h. The mixture was concentrated to give1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (88 mg, 98% yield) as yellow oil. LC-MS (ESI) found: 489 [M+H]⁺.

Step 3: To a solution of1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (1 equiv.) in DMF (2 mL) is added HATU (1.5 equiv.), DIPEA (3equiv.) andN-((3aR,4R,7S,7aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-3-(pyridin-4-yl)-1,2,4-thiadiazol-5-amine(1 equiv.) at 0° C. The mixture is stirred at rt for 2 h. The mixture isquenched with H₂O, extracted with DCM and washed with brine. The organicphase is dried and concentrated. The residual is purified by flashcolumn to giveN-(1-((3aR,4R,7S,7aR)-2,2-dimethyl-7-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11-tetraoxatridecan-13-yl)-3-((14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propenamide.

Step 4: To a solution ofN-(1-((3aR,4R,7S,7aR)-2,2-dimethyl-7-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11-tetraoxatridecan-13-yl)-3-((14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propenamide(0.01 mmol) in THF(3 mL) is added HCl (0.5 mL, 2 N in H₂O) at 0° C. Themixture is stirred at rt for 2 h. The mixture is concentrated andpurified by prep-HPLC (Method A) to giveN-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-3-((14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide(Compound 29).

Preparation of Compound 30(((S)-2-((S)-34-((1H-imidazol-5-yl)methyl)-1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-20,20-bis(1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22,32-trioxo-2,5,8,11,18,24,27,30-octaoxa-14,21,33-triazapentatriacontan-35-amido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,10H-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide)

Step 1: A solution of 2,2′-((oxybis(ethane-2, 1-diyl))bis(oxy))diaceticacid (5 g, 22.5 mmol), (4-phenylphenyl)methanol (4.2 g, 22.5 mmol) andPTSA (400 mg, 0.22 mmol) in toluene (70 mL) was stirred at 130° C. for 3h. The mixture was quenched with NaHCO₃ (aq., 10% wt.) was added andextracted with EA. The aqueous phase was extracted with EA three times,then the aqueous phase was adjusted to pH=3 with con. HCl. The aqueousphase was extracted with EA, the organic layer was concentrated to give1-([1,1′-biphenyl]-4-yl)-3-oxo-2,5,8,11-tetraoxatridecan-13-oic acid(1.4 g, 16% yield) as a colorless oil. LC-MS (ESI) found: 389 [M+H]⁺.

Step 2: A solution of1-([1,1′-biphenyl]-4-yl)-3-oxo-2,5,8,11-tetraoxatridecan-13-oic acid(1.4 g, 3.6 mmol), DIPEA (1.4 g, 10.8 mmol) and HATU (2.0 g, 5.4 mmol)in DMF(10 mL) was stirred at rt for 30 min, then di-tert-butyl3,3′-((2-amino-2-((3-(tert-butoxy)-3-oxopropoxy)methyl)propane-1,3-diyl)bis(oxy))dipropionate(2.2 g, 4.3 mmol) was added. The mixture was stirred at rt overnight,then extracted with EA. The organic layer was concentrated and theresidue was purified by flash to give [1,1′-biphenyl]-4-ylmethyl13,13-bis(((4,4-dimethyl-3-oxopentyl)oxy)methyl)-19,19-dimethyl-11,18-dioxo-3,6,9,15-tetraoxa-12-azaicosanoate(2.7 g, 86% yield) as a colorless oil. LC-MS (ESI) found: 828 [M+H]⁺.

Step 3: To a solution of [1,1′-biphenyl]-4-ylmethyl13,13-bis(((4,4-dimethyl-3-oxopentyl)oxy)methyl)-19,19-dimethyl-11,18-dioxo-3,6,9,15-tetraoxa-12-azaicosanoate(1 g, 1.1 mmol) in DCM (10 mL) was added TFA (1 mL). The mixture wasstirred at rt overnight, then purified by Prep-HPLC (Method A) to give1-([1,1′-biphenyl]-4-yl)-15,15-bis((2-carboxyethoxy)methyl)-3,13-dioxo-2,5,8,11,17-pentaoxa-14-azaicosan-20-oicacid (600 mg, 74% yield) as a colorless oil. LC-MS (ESI) found: 707[M+H]⁺.

Step 4: A solution of1-([1,1′-biphenyl]-4-yl)-15,15-bis((2-carboxyethoxy)methyl)-3,13-dioxo-2,5,8,11,17-pentaoxa-14-azaicosan-20-oicacid (70 mg, 0.1 mmol), DIPEA (38 mg, 0.3 mmol) and HATU (169 mg, 0.35mmol) in DMF (2 mL) was stirred at rt for 30 min, thenN-((3aR,4R,6R,7R,7aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(155 mg, 0.35 mmol) was added. The mixture was stirred at rt for 3 h,then extracted with EA. The organic layer was concentrated and theresidue was purified by flash followed by prep-HPLC to give[1,1′-biphenyl]-4-ylmethyl1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,24,27,30-octaoxa-14,21-diazadotriacontan-32-oate(60 mg, 30% yield) as a colorless oil.LC-MS (ESI) found: 1003 [M+2H]²⁺.

Step 5: A solution of [1,1′-biphenyl]-4-ylmethyl1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,24,27,30-octaoxa-14,21-diazadotriacontan-32-oate(60 mg, 0.03 mmol) in MeOH (4 mL) was added Pd/C (10 mg, 10% wt, 60%wet). The mixture was stirred at rt under H₂ for 2 h.

The mixture was filtered through a Celite pad, and the filtrate wasconcentrated, then purified by prep-HPLC (Method B) to give1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,24,27,30-octaoxa-14,21-diazadotriacontan-32-oicacid (40 mg, 73% yield) as a colorless oil.LC-MS (ESI) found: 1837[M+H]⁺.

Step 6: A solution of1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18,24,27,30-octaoxa-14,21-diazadotriacontan-32-oicacid (40 mg, 0.022 mmol), DIPEA (8.4 mg, 0.065 mmol) and HATU (12.4 mg,0.033 mmol) in DMF (2 mL) was stirred at rt for 30 min, then OPT-NH₂ (42mg, 0.023 mmol) was added. The mixture was stirred at rt overnight andpurified by prep-HPLC (Method B) to give(S)-2-((S)-34-((1H-imidazol-5-yl)methyl)-1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22,32-trioxo-2,5,8,11,18,24,27,30-octaoxa-14,21,33-triazapentatriacontan-35-amido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(40 mg, 50% yield) as a colorless oil. LC-MS (ESI) found: 919 [M+4H]⁺.

Step 7: To a solution of(S)-2-((S)-34-((1H-imidazol-5-yl)methyl)-1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-20,20-bis(1-((3aR,4R,6R,7R,7aR)-7-acetamido-6-methoxy-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22,32-trioxo-2,5,8,11,18,24,27,30-octaoxa-14,21,33-triazapentatriacontan-35-amido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,10H-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(20 mg, 0.005 mmol) in THE (2 mL) was added HCl (0.1 mL, 2 N in H₂O).The mixture was stirred at rt for 2 h, purified by prep-HPLC (Method A)to give((S)-2-((S)-34-((1H-imidazol-5-yl)methyl)-1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-20,20-bis(1-((2R,3R,4R,5R,6R)-5-acetamido-3,4-dihydroxy-6-methoxytetrahydro-2H-pyran-2-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22,32-trioxo-2,5,8,11,18,24,27,30-octaoxa-14,21,33-triazapentatriacontan-35-amido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(Compound 30, 8 mg, 41% yield) as a white solid. LC-MS (ESI) found: 711[M+5H]⁵.

Preparation of Compound 31((S)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)-2-((S)-2-(3-((1r,3S)-3-((20,20-bis(1-((1S,2R,3R,4R,5S)-2,3-dihydroxy-4-(2,2,2-trifluoroacetamido)-6,8-dioxabicyclo[3.2.1]octan-1-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-1-((1S,2R,3R,4R,5S)-2,3-dihydroxy-4-(2,2,2-trifluoroacetamido)-6,8-dioxabicyclo[3.2.1]octan-1-yl)-15,22-dioxo-2,5,8,11,18-pentaoxa-14,21-diazatetracosan-24-yl)oxy)cyclobutoxy)propanamido)-3-(1H-imidazol-5-yl)propanamido)pentanediamide)

Step 1: To a solution of (1r,3r)-3-(benzyloxy)cyclobutanol (1.3 g, 7.29mmol) and tetrabutylammonium chloride (0.67 g, 2.41 mmol)-in DCM (20mL). NaH (1.22 g, 30.63 mmol) in H₂O (3.5 mL) was added. The mixture wasstirred for 10 min. The reaction cooled to 0° C., tert-5 Butyl acrylate(2.8 g, 21.88 mmol) was added dropwise over 1 h. The reaction wasstirred at rt overnight and concentrated in vacuo. The crude product waspurified by flash chromatography (silica gel, 0˜10% EtOAc in PE) to givetert-butyl 3-((1r,3r)-3-(benzyloxy)cyclobutoxy)propanoate (1.6 g, 5.22mmol, 7200 yield) as a colorless oil. LC-MS (ESI) found: 329 [M+Na]. ¹HNMR (400 MHz, CDCl₃): 7.36-7.26 (m, 5H), 4.41 (d, J=4.7 Hz, 10 2H),4.27-4.11 (m, 2H), 3.56 (t, J=6.5 Hz, 2H), 2.46 (t, J=6.5 Hz, 2H),2.31-2.18 (m, 4H), 1.45 (s, 9H).

Step 2: To a solution of tert-butyl3-((1r,3r)-3-(benzyloxy)cyclobutoxy)propanoate (1.8 g, 5.88 mmol) inEtOAc (25 mL) was added Pd/C (60 mg, 1% wt, 60%0 wet) at rt under H₂(15Psi). The reaction was stirred at rt for 2 h. The resulting mixture wasfiltered and concentrated in vacuo. The 15 crude product tert-butyl3-((1r,3r)-3-hydroxycyclobutoxy)propanoate (1.1 g) was used to next stepwith no further purification. LC-MS (ESI) found: 217 [M+H]⁺.

Step 3: To a solution of tert-butyl3-((1r,3r)-3-hydroxycyclobutoxy)propanoate (1.1 g, crude) and Cs₂CO₃(3.31 g, 10.17 mmol) in DMIF (15 mL) was added methyl acrylate (1.83 mL,20.34 mmol) at rt. The mixture was stirred overnight. The resultingmixture was filtered and concentrated in vacuo. The crude product waspurified by flash chromatography (silica gel, 0˜10% EtOAc in PE) to givetert-butyl 3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanoate(400 mg, 26% yield) as a colorless oil. LC-MS (ESI) found: 303 [M+H]. ¹HNMR (400 MHz, CDCl₃):δ 4.14-4.05 (m, 2H), 3.69 (s, 3H), 3.59 (t, J=6.4Hz, 2H), 3.54 (t, J=6.5 Hz, 2H), 2.56 (t, J=6.4 Hz, 2H), 2.46 (t, J=6.5Hz, 2H), 2.20 (t, J=5.7 Hz, 4H), 1.45 (s, 9H).

Step 4: To a solution of tert-butyl3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanoate (100 mg,0.33 mmol) in DCM (3 mL) was added TFA (1 mL, 13.5 mmol) dropwise at 0°C. under N₂. The reaction was stirred at rt for 1 h. The resultingmixture was filtered and concentrated in vacuo. The crude product3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy) propanoic acid (80 mg)was used to next step with no further purification. LC-MS (ESI) found:247 [M+H]⁺.

Step 5: To a solution of3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanoic acid (200 mg,0.81 mmol) and HATU (401 mg, 1.1 mmol) in DMF (5 mL) was stirred at rtfor 30 min, and then added di-tert-butyl3,3′-((2-amino-2-((3-(tert-butoxy)-3-oxopropoxy)methyl)propane-1,3-diyl)bis(oxy))dipropanoate(492.8 mg, 0.97 mmol) and N,N-Diisopropylethylamine(0.4 mL, 2.4 mmol) atrt. The reaction was stirred overnight. The resulting mixture wasdiluted with DCM (100 mL), washed with H₂O (50 mL×2) and brine (50 mL),dried over Na₂SO₄. The organic layer was separated and concentrated invacuo. The crude product was purified by flash chromatography (silicagel, 0˜80% EA in PE) to give di-tert-butyl3,3′-((2-((3-(tert-butoxy)-3-oxopropoxy)methyl)-2-(3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanamido)propane-1,3-diyl)bis(oxy))dipropanoate(300 mg, 50% yield) as a colorless oil. LC-MS (ESI) found: 734 [M+H]⁺.

Step 6: To a solution of di-tert-butyl3,3′-((2-((3-(tert-butoxy)-3-oxopropoxy)methyl)-2-(3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanamido)propane-1,3-diyl)bis(oxy))dipropanoate(300 mg, 0.41 mmol) in DCM (4 mL) was added TFA (1 mL, 13.5 mmol)dropwise at 0° C. under N₂. The reaction was stirred at rt for 2 h. Theresulting mixture was filtered and concentrated in vacuo. The crudeproduct was purified by prep-HPLC (Method A) to give3,3′-((2-((2-carboxyethoxy)methyl)-2-(3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanamido)propane-1,3-diyl)bis(oxy))dipropanoicacid (200 mg, 86%) as a colorless oil. LC-MS (ESI) found: 566 [M+H]⁺. ¹HNMR (400 MHz, CDCl₃): δ 6.52 (s, 1H), 4.13 (dd, J=11.1, 6.0 Hz, 2H),3.71 (dd, J=7.6, 3.8 Hz, 15H), 3.60 (t, J=6.3 Hz, 2H), 3.56 (t, J=5.9Hz, 2H), 2.57 (t, J=6.0 Hz, 8H), 2.43 (t, J=5.8 Hz, 2H), 2.22 (t, J=5.7Hz, 4H).

Step 7: A solution of3,3′-((2-((2-carboxyethoxy)methyl)-2-(3-((1r,3r)-3-(3-methoxy-3-oxopropoxy)cyclobutoxy)propanamido)propane-1,3-diyl)bis(oxy))dipropanoicacid (15 mg, 0.03 mmol) and HATU (35.9 mg, 0.09 mmol) in DMF (1 mL) wasstirred at rt for 30 min, thenN-((3aR,4S,7S,8R,8aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-2,2-dimethylhexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-8-yl)-2,2,2-trifluoroacetamide(47.5 mg, 0.09 mmol) and N,N-Diisopropylethylamine (0.02 mL, 0.14 mmol)were added at rt. The reaction was stirred for 2.5 h.

The residue was purified by prep-HPLC (Method A) to give methyl 3-((1r,3r)-3-((1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-20,20-bis(1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18-pentaoxa-14,21-diazatetracosan-24-yl)oxy)cyclobutoxy)propanoate(6 mg, 11% yield) as a colorless oil. LC-MS (ESI) found: 1010 [M+2]²⁺.¹H NMR (400 MHz, MeOD): δ 5.29 (d, J=1.7 Hz, 3H), 4.35 (q, J=5.9 Hz,6H), 4.10 (dd, J=9.8, 5.6 Hz, 2H), 3.97-3.88 (m, 9H), 3.80 (dd, J=9.0,4.6 Hz, 6H), 3.71-3.61 (m, 53H), 3.57-3.53 (m, 8H), 3.38 (t, J=5.5 Hz,6H), 2.55 (t, J=6.1 Hz, 2H), 2.44 (q, J=6.1 Hz, 8H), 2.19 (t, J=5.6 Hz,4H), 1.50 (s, 9H), 1.35 (s, 9H).

Step 8: To a solution of methyl3-((1r,3r)-3-((1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-20,20-bis(1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18-pentaoxa-14,21-diazatetracosan-24-yl)oxy)cyclobutoxy)propanoate(6 mg) in DCM (2 mL) was added Me₃SnOH (0.45 mg, 0.002 mmol) at rt underN₂. The reaction was stirred at 80° C. for 3 days. The resulting mixturewas filtered and concentrated in vacuo. The crude product3-((1r,3r)-3-((1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-20,20-bis(1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18-pentaoxa-14,21-diazatetracosan-24-yl)oxy)cyclobutoxy)propanoicacid (3 mg) was used to next step with no further purification. LC-MS(ESI) found: 1003 [M+2H]²⁺.

Step 9: A solution of3-((1r,3r)-3-((1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-20,20-bis(1-((3aR,4S,7S,8R,8aR)-2,2-dimethyl-8-(2,2,2-trifluoroacetamido)hexahydro-4,7-epoxy[1,3]dioxolo[4,5-d]oxepin-4-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-15,22-dioxo-2,5,8,11,18-pentaoxa-14,21-diazatetracosan-24-yl)oxy)cyclobutoxy)propanoicacid (3 mg, 0.001 mmol) and HATU (0.85 mg, 0.002 mmol) in DMF(1 mL) wasstirred at rt for 30 min, then OPT-NH₂ (3.05 mg, 0.002 mmol) andN,N-Diisopropylethylamine (0.05 mL) were added at rt. The reaction wasstirred for 2.5 h. The residue was purified by prep-HPLC (Method A) togive target (1 mg) as a colorless oil. LC-MS (ESI) found: 1280 [M+3H]³⁺

Step 10: To a solution of product from Step 9 (1 mg) in THE (3 mL) wasadded HCl (0.1 mL, 1 M in H₂O). The reaction was stirred at rt for 3 h.The crude product was purified by prep-HPLC (Method A) to give(S)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)-2-((S)-2-(3-((1r,3S)-3-((20,20-bis(1-((1S,2R,3R,4R,5S)-2,3-dihydroxy-4-(2,2,2-trifluoroacetamido)-6,8-dioxabicyclo[3.2.1]octan-1-yl)-15-oxo-2,5,8,11,18-pentaoxa-14-azanonadecan-19-yl)-1-((1S,2R,3R,4R,5S)-2,3-dihydroxy-4-(2,2,2-trifluoroacetamido)-6,8-dioxabicyclo[3.2.1]octan-1-yl)-15,22-dioxo-2,5,8,11,18-pentaoxa-14,21-diazatetracosan-24-yl)oxy)cyclobutoxy)propanamido)-3-(1H-imidazol-5-yl)propanamido)pentanediamide(Compound 31, 0.9 mg) as a colorless oil. LC-MS (ESI) found: 1240[M+3H]3.

Preparation of Compound 32:3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((2R,3R,4R,5R)-5-((2,6-dimethoxypyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)

Step 1: To a solution of((3aR,4R,7R,7aR)-7-((2,6-dimethoxypyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanolin DMF is added NaH at 0° C. and stirred for 1 h under N₂. Then2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl 4-methylbenzenesulfonate isadded at 0° C. and stirred at rt for 2 h under N₂. The resulting mixtureis diluted with DCM and water. The aqueous phase is extracted with DCM.The organic layer is separated, dried over Na₂SO₄ and concentrated invacuo. The crude product is purified by flash chromatography (silicagel, 0-10% MeOH in DCM) to giveN-((3aR,4R,7R,7aR)-4-(13-azido-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-2,6-dimethoxypyrimidin-4-amine.

Step 2: To a solutionN-((3aR,4R,7R,7aR)-4-(13-azido-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-2,6-dimethoxypyrimidin-4-aminein MeOH is added Pd/C at rt under a H₂ balloon. The reaction is stirredat rt for 1.5 h. The resulting mixture is filtered and concentrated invacuo. The crude productN-((3aR,4R,7R,7aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-2,6-dimethoxypyrimidin-4-amineis used to next step with no further purification.

Step 3: To a solution of13-((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid and HATU in DMF is stirred at rt for 30 min, thenN-((3aR,4R,7R,7aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-2,6-dimethoxypyrimidin-4-amineand N,N-Diisopropylethylamine are added at rt. The reaction is stirredovernight. The residue is purified by prep-HPLC (Method A) to give3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((3aR,4R,7R,7aR)-7-((2,6-dimethoxypyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)as a yellow oil.

Step 4: To a solution of3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((3aR,4R,7R,7aR)-7-((2,6-dimethoxypyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)in THE is added HCl (1 M in H₂O).The reaction is stirred at rt for 3 h.The crude product is purified by prep-HPLC (Method A) to give3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(1-((2R,3R,4R,5R)-5-((2,6-dimethoxypyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(Compound 32).

Preparation of Compound 333,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)tris(oxy))tris(N-(1-((2R,3R,4R,5R)-5-acetamido-3,4-dihydroxytetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)

Step 1: To a solution of13,13-bis((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (10 mg, 0.014 mmol) and HATU (19.2 mg, 0.05 mmol) in DMF (2 mL) wasstirred at rt for 30 min, thenN-((3aR,4R,7R,7aR)-4-(13-amino-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-3aH-[1,3]dioxolo[4,5-c]pyran-7-yl)acetamide(24.3 mg, 0.06 mmol) and DIPEA (0.01 mL, 0.07 mmol) were added at rt.The reaction was stirred overnight. The residue was purified byprep-HPLC (Method A) to give product (10 mg, 37% yield) as yellow oil.LC-MS (ESI) found: 1900 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 9.13 (d, J=2.6Hz, 1H), 8.81 (s, 1H), 8.27 (dd, J=9.5, 2.6 Hz, 1H), 8.08 (s, 1H),7.00-6.97 (m, 3H), 6.84 (d, J=7.6 Hz, 1H), 5.91 (d, J=8.5 Hz, 3H), 4.39(d, J=5.8 Hz, 6H), 4.26 (dd, J=7.1, 2.0 Hz, 3H), 3.92-3.81 (m, 9H),3.72-3.68 (m, 28H), 3.64 (dd, J=8.6, 4.3 Hz, 40H), 3.55 (t, J=5.3 Hz,6H), 3.45-3.40 (m, 6H), 2.43 (t, J=5.7 Hz, 6H), 2.01 (s, 9H), 1.50 (s,9H), 1.34 (s, 9H).

Step 2: To a solution of material from step 1 (10 mg, 0.005 mmol) in THE(3 mL) was added HCl (0.5 mL, 1 M in H₂O). The reaction was stirred atrt for 3 h. The crude product was purified by prep-HPLC (Method A) togive3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)tris(oxy))tris(N-(1-((2R,3R,4R,5R)-5-acetamido-3,4-dihydroxytetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)propanamide)(Compound 33, 0.7 mg, 7.5% yield) as a yellow oil. LC-MS (ESI) found:1780 [M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 9.05 (d, J=2.7 Hz, 1H), 8.31 (dd,J=9.6, 2.7 Hz, 1H), 7.25 (d, J=9.7 Hz, 1H), 4.07 (s, 3H), 3.91-3.86 (m,8H), 3.82 (t, J=5.2 Hz, 3H), 3.76-3.74 (m, 3H), 3.71-3.66 (m, 24H),3.65-3.62 (m, 36H), 3.57-3.47 (m, 15H), 3.37 (t, J=5.5 Hz, 6H), 2.44 (t,J=6.1 Hz, 6H), 2.00 (s, 9H).

Preparation of Compound 34:3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-((1-(11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-1H-1,2,3-triazol-4-yl)methyl)propanamide)

Step 1: To a solution of(2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-2-(hydroxymethyl)tetrahydro-2H-pyran-3,4-diol(1 equiv.) in 2,2-dimethoxypropane is added TsOH (0.1 equiv.), and thereaction is stirred at room temperature overnight. The 5 mixture isconcentrated under reduced pressure to give a crude product((3aR,4R,7S,7aR)-7-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanol.

Step 2: To a solution of((3aR,4R,7S,7aR)-7-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanolin DMF at 0° C. is 10 added NaH (600% wt. in mineral oil) and 1-azido-1l-bromoundecane, then the mixture is stirred at the room temperatureovernight. On consumption of starting material (LCMS monitoring), thereaction vessel is again cooled to 0° C., water slowly added, and thereaction mixture is stirred for 15 min. The mixture is diluted withethyl acetate and washed with water. The organic layer is dried oversodium sulfate, concentrated under reduced pressure to give a crudeproduct, which is 15 purified by flash chromatography to giveN-((3aR,4R,7S,7aR)-4-(((1I-azidoundecyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-6-chloro-2-(trifluoromethyl)pyrimidin-4-amine.

Step 3: To a solution of13-((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (1 equiv.) in DMIF (2 mL) is added HATU (1.5 20 equiv.), DIPEA (3equiv.) and propargylamine (2 equiv.) at 0° C. The mixture is stirred atrt for 2 h. The mixture is quenched with H₂O, extracted with DCM andwashed with brine. The organic phase is dried and concentrated. Theresidual is purified by flash column to give3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(prop-2-yn-1-yl)propanamide).

Step 4: THPTA and CuSO₄ are dissolved in water. The mixture is added toa solution ofN-((3aR,4R,7S,7aR)-4-(((11-azidoundecyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-6-chloro-2-(trifluoromethyl)pyrimidin-4-amineand3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(prop-2-yn-1-yl)propanamide)in MeOH. A solution of Na ascorbate in water is added. The reaction isstirred overnight. The resulting mixture is filtered. The crude productis purified by prep-HPLC (Method A) to give3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-((1-(11-(((3aR,4R,7S,7aR)-7-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methoxy)undecyl)-1H-1,2,3-triazol-4-yl)methyl)propanamide).

Step 5: To a solution of3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-((1-(11-(((3aR,4R,7S,7aR)-7-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methoxy)undecyl)-1H-1,2,3-triazol-4-yl)methyl)propanamide)in THE is added HCl (2N in H₂O) at 0° C. The mixture is stirred at rtfor 2 h. The mixture is concentrated and purified by prep-HPLC (MethodA) to give3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-((1-(11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-1H-1,2,3-triazol-4-yl)methyl)propanamide)(Compound 34).

Preparation of Compound 35((2R,3R,4R,5S)-2-(13-((2,4-dinitrophenyl)amino)-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol)

Step 1: To a solution of(2R,3R,4R,5S)-2-(13-amino-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(72 mg, 0.149 mmol) in DMF (3 mL) was added K₂CO₃ (62 mg, 0.446 mmol)and 1-chloro-2,4-dinitrobenzene (30 mg, 0.149 mmol). The mixture wasstirred at 80° C. for 16 h. The mixture was filtered, concentrated andpurified by prep-HPLC (Method A) to give(2R,3R,4R,5S)-2-(13-((2,4-dinitrophenyl)amino)-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(Compound 35, 7.0 mg, 7% yield) as yellow solid. LC-MS (ESI) found: 651[M+H]⁺. ¹H NMR (400 MHz, MeOD): δ 9.02 (d, J=2.7 Hz, 1H), 8.50 (d, J=4.8Hz, 1H), 8.28 (dd, J=9.6, 2.7 Hz, 1H), 7.22 (d, J=9.6 Hz, 1H), 6.89 (d,J=4.9 Hz, 1H), 4.34 (td, J=10.5, 5.2 Hz, 1H), 4.07 (dd, J=10.9, 5.2 Hz,1H), 3.90 (d, J=2.9 Hz, 1H), 3.81 (t, J=5.2 Hz, 2H), 3.71-3.61 (m, 18H),3.15 (t, J=10.9 Hz, 1H).

Preparation of Compound 36:N-(11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((2-((3-((2-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide

Step 1: To a solution ofN-((3aR,4R,7S,7aR)-4-(((11-azidoundecyl)oxy)methyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-6-chloro-2-(trifluoromethyl)pyrimidin-4-aminein THE is added PPh₃ and water. The reaction mixture is stirred at rtovernight. The mixture is concentrated in vacuo. The crude product ispurified by flash chromatography (silica gel, 0-10% MeOH in DCM) to give(2R,3R,4R,5S)-2-(((11-aminoundecyl)oxy)methyl)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol.

Step 2: To a solution of13-((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (1 equiv.) in DMF (2 mL) is added HATU (1.5 equiv.), DIPEA (3equiv.) and(2R,3R,4R,5S)-2-(((11-aminoundecyl)oxy)methyl)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(2 equiv.) at 0° C. The mixture is stirred at rt for 2 h. The mixture isquenched with H₂O, extracted with DCM and washed with brine. The organicphase is dried and concentrated. The residual is purified by flashcolumn to giveN-(11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((2-((3-((2-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propenamide(Compound 36).

Preparation of Compound 37:3,3′-((2-((3-((11-(((2R,3R,4R,5R)-5-((1,2,4-thiadiazol-5-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(11-(((2R,3R,4R,5R)-5-((1,2,4-thiadiazol-5-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)propanamide)

Step 1: To a solution of13,13-bis((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (1 equiv.) in DMF (2 mL) is added HATU (1.5 equiv.), DIPEA (3equiv.) and(2R,3R,4R,5R)-5-((1,2,4-thiadiazol-5-yl)amino)-2-(((11-aminoundecyl)oxy)methyl)tetrahydro-2H-pyran-3,4-diol(3 equiv.) at 0° C. The mixture is stirred at rt for 2 h. The mixture isquenched with H₂O, extracted with DCM and washed with brine.

The organic phase is dried and concentrated. The residual is purified byflash column to give3,3′-((2-((3-((11-(((2R,3R,4R,5R)-5-((1,2,4-thiadiazol-5-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-(11-(((2R,3R,4R,5R)-5-((1,2,4-thiadiazol-5-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)propanamide)(Compound 37).

Preparation of Compound 38:N-(2-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide

Step 1: To a solution of1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (1 equiv.) in DMF (2 mL) is added HATU (1.5 equiv.), DIPEA (3equiv.) and(2R,3R,4R,5S)-2-(((11-aminoundecyl)oxy)methyl)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(1 equiv.) at 0° C. The mixture is stirred at rt for 2 h. The mixture isquenched with H₂O, extracted with DCM and washed with brine. The organicphase is dried and concentrated. The residual is purified by flashcolumn to giveN-(2-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide(Compound 38).

Preparation of Compound 39(S)-2-((S)-2-((1H-imidazol-5-yl)methyl)-23-(1-(1-((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)-20-((3-((11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-15,22-dioxo-2,18-dioxa-14,21-diazaheptacosan-27-yl)-1H-1,2,3-triazol-4-yl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazatricosanamido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,10H-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide

Step 1: To a solution of3,3′-((2-(6-azidohexanamido)propane-1,3-diyl)bis(oxy))dipropionic acid(1 equiv.) in DMF (2 mL) is added HATU (1.5 equiv.), DIPEA (3 equiv.)and(2R,3R,4R,5S)-2-(((11-aminoundecyl)oxy)methyl)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)tetrahydro-2H-pyran-3,4-diol(2 equiv.) at 0° C. The mixture is stirred at rt for 2 h. The mixture isquenched with H₂O, extracted with DCM and washed with brine. The organicphase is dried and concentrated. The residual is purified by flashcolumn to giveN-(11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((2-((3-((2-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propenamide.

Step 2: THPTA and CuSO₄ are dissolved in water. The mixture is added toa solution ofN-(11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((2-((3-((2-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propenamideand OPT-Alkyne in MeOH. A solution of Na ascorbate in water is added.The reaction is stirred overnight. The resulting mixture is filtered.

The crude product is purified by prep-HPLC (Method A) to give(S)-2-((S)-2-((1H-imidazol-5-yl)methyl)-23-(1-(1-((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)-20-((3-((11-(((2R,3R,4R,5S)-5-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-15,22-dioxo-2,18-dioxa-14,21-diazaheptacosan-27-yl)-1H-1,2,3-triazol-4-yl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazatricosanamido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(Compound 39).

Preparation of Compound 48:(2R,3R,4R,5S)-5-((3-bromo-1,2,4-thiadiazol-5-yl)amino)-2-(13-((2,4-dinitrophenyl)amino)-2,5,8,11-tetraoxatridecyl)tetrahydro-2H-pyran-3,4-diol

Step 1: NaH is added to a suspension of((3aR,4R,7S,7aR)-7-((3-bromo-1,2,4-thiadiazol-5-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanolin dry DMF. The mixture is stirred at rt for 1.5 h. Then2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)ethyl4-methylbenzenesulfonate is added to the above solution. The mixture isfurther stirred at rt for 16 h. To the mixture is added NH₄Cl (aq) andthe solvent removed. Then the residual is purified by flash(DCM:MeOH=15:1) to give3-bromo-N-((3aR,4R,7S,7aR)-4-(13-((2,4-dinitrophenyl)amino)-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)-1,2,4-thiadiazol-5-amine.

Step 2: To a solution of3,3′-((2-(2-(2-(2-(2-((2,4-dinitrophenyl)amino)ethoxy)ethoxy)ethoxy)acetamido)propane-1,3-diyl)bis(oxy))bis(N-((1-(11-(((3aR,4R,7S,7aR)-7-((6-chloro-2-(trifluoromethyl)pyrimidin-4-yl)amino)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methoxy)undecyl)-1H-1,2,3-triazol-4-yl)methyl)propanamide)in THE is added HCl (2N in H₂O) at 0° C. The mixture is stirred at rtfor 2 h. The mixture is concentrated and purified by prep-HPLC (MethodA) to give(2R,3R,4R,5S)-5-((3-bromo-1,2,4-thiadiazol-5-yl)amino)-2-(13-((2,4-dinitrophenyl)amino)-2,5,8,11-tetraoxatridecyl)tetrahydro-2H-pyran-3,4-diol(Compound 48).

Preparation of Compound 49:(2R,3R,4R,5S)-5-((3-bromo-1,2,4-thiadiazol-5-yl)amino)-2-(22-((2,4-dinitrophenyl)amino)-2,5,8,11,14,17,20-heptaoxadocosyl)tetrahydro-2H-pyran-3,4-diol

It is prepared according to the procedures same as that for Compound 48by using 20-((2,4-dinitrophenyl)amino)-3,6,9,12,15,18-hexaoxaicosyl4-methylbenzenesulfonate as the starting material.

Preparation of Compound 50:(2R,3R,4R,5S)-2-(22-((2,4-dinitrophenyl)amino)-2,5,8,11,14,17,20-heptaoxadocosyl)-5-((3-(pyridin-4-yl)-1,2,4-thiadiazol-5-yl)amino)tetrahydro-2H-pyran-3,4-diol

It is prepared according to the procedures same as that for A357 byusing Compound 49 as the starting material.

Preparation of Compound 51(S)-2-((S)-2-((1H-imidazol-5-yl)methyl)-23-(1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazatricosanamido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,10H-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide

Step 1: THPTA (2.97 mg, 0.007 mmol) and Cu₂SO₄ (0.22 mg, 0.001 mmol)were dissolved in water (0.5 mL). Then a solution of(2R,3R,4R,5S)-2-(13-azido-2,5,8,11-tetraoxatridecyl)-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(A419, 7.69 mg, 0.015 mmol) and OPT-Alkyne (30 mg, 0.014 mmol) in MeOH(2 mL) was added to the above mixture.

Then a freshly-prepared solution of Na ascorbate (0.54 mg, 0.003 mmol)in water (0.5 mL) was added and the reaction mixture was stirred at roomtemperature for 24 hours. The mixture was filtered and the residue waspurified by pre-HPLC (Method A) to afford(S)-2-((S)-2-((1H-imidazol-5-yl)methyl)-23-(1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazatricosanamido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(14 mg, 36% yield). LC-MS (ESI) found: 676 [M+4H]⁺. ¹H NMR (400 MHz,D₂O): δ8.43 (d, J=4.6 Hz, 1H), 8.37 (s, 1H), 7.74 (s, 1H), 7.08 (d,J=8.3 Hz, 2H), 6.98 (s, 1H), 6.93-6.84 (m, 4H), 6.83-6.73 (m, 4H),4.52-4.44 (m, 5H), 4.40-4.31 (m, 4H), 4.28-4.18 (m, 3H), 4.15-4.04 (m,4H), 3.97 (dd, J=10.9, 5.0 Hz, 2H), 3.91 (d, J=3.2 Hz, 2H), 3.79 (ddd,J=30.0, 10.0, 4.0 Hz, 1OH), 3.56 (ddd, J=19.4, 12.8, 7.5 Hz, 46H), 3.28(d, J=5.4 Hz, 3H), 3.17 (dd, J=14.4, 8.8 Hz, 5H), 3.07-2.95 (m, 4H),2.83 (d, J=38.1 Hz, 8H), 2.63 (dd, J=17.2, 9.6 Hz, 6H), 2.45 (dd,J=10.1, 5.2 Hz, 3H), 2.19 (t, J=7.3 Hz, 4H), 2.14-2.00 (m, 6H),1.94-1.76 (m, 11H), 1.59 (dd, J=7.7, 2.6 Hz, 6H), 1.40 (d, J=10.1 Hz,2H), 1.11-0.97 (m, 5H), 0.88-0.52 (m, 24H).

Preparation of Compound 52(S)-2-((S)-2-((1H-imidazol-5-yl)methyl)-23-(1-(6-((1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)amino)-6-oxohexyl)-1H-1,2,3-triazol-4-yl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazatricosanamido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,10H-pyrrolo[2,1

Step 1: THPTA (3.91 mg, 0.009 mmol) and Cu₂SO₄ (0.29 mg, 0.002 mmol)were dissolved in water (0.5 mL). Then a solution of6-azido-N-(1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)hexanamide(A417, 25.96 mg, 0.020 mmol) and OPT-Alkyne (39 mg, 0.018 mmol) in MeOH(3 mL) was added into the above mixture. A freshly-prepared solution ofNa ascorbate (0.71 mg, 0.004 mmol) in water (0.5 mL) was then added andthe reaction mixture was stirred at room temperature for 24 hours.Solvent was evaporated and the crude material was purified by pre-HPLC(Method A) to afford(S)-2-((S)-2-((1H-imidazol-5-yl)methyl)-23-(1-(6-((1,3-bis((1-(1-((2R,3R,4R,5S)-3,4-dihydroxy-5-((4-(trifluoromethyl)pyrimidin-2-yl)amino)tetrahydro-2H-pyran-2-yl)-2,5,8,11-tetraoxatridecan-13-yl)-1H-1,2,3-triazol-4-yl)methoxy)propan-2-yl)amino)-6-oxohexyl)-1H-1,2,3-triazol-4-yl)-4,20-dioxo-7,10,13,16-tetraoxa-3,19-diazatricosanamido)-N1-((S)-1-(((3S,6R,11R,14S,17S,20S,23S,29S,34aS)-6-(((S)-1-(((S)-1-(((2S,3R)-1-amino-3-hydroxy-1-oxobutan-2-yl)amino)-3-hydroxy-1-oxopropan-2-yl)amino)-1-oxo-3-phenylpropan-2-yl)carbamoyl)-23,29-bis(3-guanidinopropyl)-20-(4-hydroxybenzyl)-17-(hydroxymethyl)-14-isobutyl-3-isopropyl-1,4,12,15,18,21,24,27,30-nonaoxotriacontahydro-1H,1OH-pyrrolo[2,1-j][1,2]dithia[5,8,11,14,17,20,23,26,29]nonaazacyclodotriacontin-11-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(10 mg, 17% yield). LC-MS (ESI) found: 705 [M+5H]”. ¹H NMR (400 MHz,MeOD): δ 8.79 (s, 1H), 8.50 (d, J=4.9 Hz, 2H), 8.03 (s, 2H), 7.78 (s,1H), 7.36 (s, 1H), 7.17-7.02 (m, 7H), 6.89 (d, J=4.9 Hz, 2H), 6.74 (d,J=8.4 Hz, 2H), 5.78 (dt, J=22.6, 11.6 Hz, 2H), 5.24-5.10 (m, 3H), 5.05(d, J=4.7 Hz, 2H), 4.79-4.69 (m, 2H), 4.57 (d, J=6.9 Hz, 8H), 4.52 (d,J=2.7 Hz, 1H), 4.43-4.29 (m, 7H), 4.19 (dd, J=10.4, 4.8 Hz, 2H), 4.06(dd, J=5 10.9, 5.1 Hz, 2H), 3.89 (dd, J=10.0, 4.3 Hz, 7H), 3.78-3.47 (m,59H), 3.34 (d, J=5.6 Hz, 3H), 3.23 (t, J=7.1 Hz, 2H), 3.14 (t, J=10.9Hz, 4H), 3.06-2.88 (m, 6H), 2.81 (dd, J=17.6, 9.7 Hz, 2H), 2.70 (dd,J=17.5, 8.4 Hz, 3H), 2.61-2.34 (m, 3H), 2.22 (dt, J=21.7, 7.3 Hz, 7H),2.09-1.80 (m, 14H), 1.77-1.69 (m, 4H), 1.69-1.52 (m, 4H), 1.39-1.13 (m,7H), 1.06-0.93 (m, 10H), 0.85 (t, J=6.2 Hz, 9H).

Preparation of Compound 533-(2-(2-(2-(3-(1-(4-amino-6,7-dimethoxyquinazolin-2-yl)-4-((R)-3-(ethylamino)-3-(4-fluorophenyl)propanoyl)piperazin-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)-N-(1,31-bis((2R,3R,4R,5R)-3,4-dihydroxy-5-(2-oxooxazolidin-3-yl)tetrahydro-2H-pyran-2-yl)-16-(15-((2R,3R,4R,5R)-3,4-dihydroxy-5-(2-oxooxazolidin-3-yl)tetrahydro-2H-pyran-2-yl)-2,5,8,11,14-pentaoxapentadecyl)-2,5,8,11,14,18,21,24,27,30-decaoxahentriacontan-16-yl)propanamide

Step 1: To a solution of3-((3R,4R,5R,6R)-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-3-yl)oxazolidin-2-one(1 equiv.) in 2,2-dimethoxypropane is added TsOH (0.1 equiv.), and thereaction is stirred at room temperature overnight. The mixture isconcentrated under reduced pressure to give a crude product3-((3aR,4R,7R,7aR)-4-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)oxazolidin-2-one.

Step 2: To a solution of3-((3aR,4R,7R,7aR)-4-(hydroxymethyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)oxazolidin-2-onein DMF at 0° C. is added NaH (60% wt. in mineral oil) and1-bromo-2-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethane, then the mixture isstirred at the room temperature overnight. On consumption of startingmaterial (LCMS monitoring), the reaction vessel is again cooled to 0°C., water slowly added, and the reaction mixture is stirred for 15 min.The mixture is diluted with ethyl acetate and washed with water.

The organic layer is dried over sodium sulfate, concentrated underreduced pressure to give a crude product, which is purified by flashchromatography to give3-((3aR,4R,7R,7aR)-4-(13-bromo-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)oxazolidin-2-one.

Step 3: To a solution of tert-butyl(tert-butoxycarbonyl)(1,3-dihydroxy-2-(hydroxymethyl)propan-2-yl)carbamatein DMF is added NaH at 0° C. and stirred for 1 h under N₂. Then3-((3aR,4R,7R,7aR)-4-(13-bromo-2,5,8,11-tetraoxatridecyl)-2,2-dimethyltetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-7-yl)oxazolidin-2-oneis added at 0° C. and stirred at rt for 2 h under N₂. The resultingmixture is diluted with DCM and water. The aqueous phase is extractedwith DCM. The organic layer is separated, dried over Na₂SO₄ andconcentrated in vacuo. The crude product is purified by flashchromatography (silica gel, 0˜10% MeOH in DCM) to give tert-butyl(1,31-bis((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-16-(15-((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11,14-pentaoxapentadecyl)-2,5,8,11,14,18,21,24,27,30-decaoxahentriacontan-16-yl)(tert-butoxycarbonyl)carbamate.

Step 4: To a solution of tert-butyl(1,31-bis((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-16-(15-((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11,14-pentaoxapentadecyl)-2,5,8,11,14,18,21,24,27,30-decaoxahentriacontan-16-yl)(tert-butoxycarbonyl)carbamatein DCM is added TFA at rt under N₂. After stirring for 2 h, the mixtureis quenched with NaHCO₃(aq) and extracted with EA. The combined organiclayer is washed with brine, dried over anhydrous Na₂SO₄. The residue isconcentrated and purified by chromatography on silica gel (0-50% ethylacetate in petroleum) to give 53-4.

Step 5: To a solution of tert-butyl((1R)-3-(4-(4-amino-6,7-dimethoxyquinazolin-2-yl)-3-(3-hydroxyphenyl)piperazin-1-yl)-1-(4-fluorophenyl)-3-oxopropyl)(ethyl)carbamatein DMF at 0° C. is added NaH (60% wt. in mineral oil) and tert-butyl3-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)propanoate, then the mixture isstirred at the room temperature overnight. On consumption of startingmaterial (LCMS monitoring), the reaction vessel is again cooled to 0°C., water slowly added, and the reaction mixture is stirred for 15 min.The mixture is diluted with ethyl acetate and washed with water. Theorganic layer is dried over sodium sulfate, concentrated under reducedpressure to give a crude product, which is purified by flashchromatography to give tert-butyl3-(2-(2-(2-(3-(1-(4-amino-6,7-dimethoxyquinazolin-2-yl)-4-((R)-3-((tert-butoxycarbonyl)(ethyl)amino)-3-(4-fluorophenyl)propanoyl)piperazin-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)propanoate.

Step 6: To a solution of tert-butyl3-(2-(2-(2-(3-(1-(4-amino-6,7-dimethoxyquinazolin-2-yl)-4-((R)-3-((tert-butoxycarbonyl)(ethyl)amino)-3-(4-fluorophenyl)propanoyl)piperazin-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)propanoatein THE at room temperature is added LiGH (5 M in water) and stirredovernight. On consumption of starting material, the mixture is dilutedwith ethyl acetate and washed with water. The organic layer is driedover sodium sulfate, concentrated under reduced pressure to give a crudeproduct, which is purified by flash chromatography to give3-(2-(2-(2-(3-(1-(4-amino-6,7-dimethoxyquinazolin-2-yl)-4-((R)-3-((tert-butoxycarbonyl)(ethyl)amino)-3-(4-fluorophenyl)propanoyl)piperazin-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)propanoicacid.

Step 7: To a solution of3-(2-(2-(2-(3-(1-(4-amino-6,7-dimethoxyquinazolin-2-yl)-4-((R)-3-((tert-butoxycarbonyl)(ethyl)amino)-3-(4-fluorophenyl)propanoyl)piperazin-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)propanoicacid (1 equiv.) in DMF is added HATU (1.5 equiv.), DIPEA (3 equiv.) and53-4 (1 equiv.) at 0° C. The mixture is stirred at rt for 2 h. Themixture is quenched with H₂O, extracted with DCM and washed with brine.The organic phase is dried and concentrated. The residual is purified byflash column to give tert-butyl((1R)-3-(4-(4-amino-6,7-dimethoxyquinazolin-2-yl)-3-(3-((1-((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-16,16-bis(15-((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11,14-pentaoxapentadecyl)-18-oxo-2,5,8,11,14,21,24,27-octaoxa-17-azanonacosan-29-yl)oxy)phenyl)piperazin-1-yl)-1-(4-fluorophenyl)-3-oxopropyl)(ethyl)carbamate.

Step 8: To a solution of tert-butyl((1R)-3-(4-(4-amino-6,7-dimethoxyquinazolin-2-yl)-3-(3-((1-((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-16,16-bis(15-((3aR,4R,7R,7aR)-2,2-dimethyl-7-(2-oxooxazolidin-3-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)-2,5,8,11,14-pentaoxapentadecyl)-18-oxo-2,5,8,11,14,21,24,27-octaoxa-17-azanonacosan-29-yl)oxy)phenyl)piperazin-1-yl)-1-(4-fluorophenyl)-3-oxopropyl)(ethyl)carbamatein THE is added HCl (2N in H₂O) at 0° C. The mixture is stirred at rtfor 2 h. The mixture is concentrated and purified by prep-HPLC (MethodA) to give3-(2-(2-(2-(3-(1-(4-amino-6,7-dimethoxyquinazolin-2-yl)-4-((R)-3-(ethylamino)-3-(4-fluorophenyl)propanoyl)piperazin-2-yl)phenoxy)ethoxy)ethoxy)ethoxy)-N-(1,31-bis((2R,3R,4R,5R)-3,4-dihydroxy-5-(2-oxooxazolidin-3-yl)tetrahydro-2H-pyran-2-yl)-16-(15-((2R,3R,4R,5R)-3,4-dihydroxy-5-(2-oxooxazolidin-3-yl)tetrahydro-2H-pyran-2-yl)-2,5,8,11,14-pentaoxapentadecyl)-2,5,8,11,14,18,21,24,27,30-decaoxahentriacontan-16-yl)propanamide(Compound 53).

Preparation of Compound 54:(2S,4R)-1-(2-(3-acetyl-5-(2-((2-((2-((2-((((2R,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)pyrimidin-5-yl)-1H-indazol-1-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide

Step 1: To a solution((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanol(1.0 equiv.), PPh₃ (1.5 equiv.) and phthalimide (1.0 eq) in dry DCM isadded DIAD (1.2 eq) dropwise at ice-bath under N₂ atmosphere. Then thereaction is allowed to warm to rt. The resulting reaction mixture isstirred at the same temperature until starting material is consumed. Themixture is evaporated. The crude product is further purified by silicagel column chromatography to give2-(((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)isoindoline-1,3-dione.

Step 2: To a solution of2-(((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)isoindoline-1,3-dionein DMF is added hydrazine (35% wt. in H₂O) and stirred with heating asnecessary overnight. On consumption of starting material (LCMSmonitoring), the reaction vessel is again cooled to room temperature.The mixture is diluted with ethyl acetate and washed with water. Theorganic layer is dried over sodium sulfate, concentrated under reducedpressure to give a crude product, which is purified by flashchromatography to give((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanamine.

Step 3: To a solution of((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanamine(1 equiv.) in DMF is added HATU (1.5 equiv.), DIPEA (3 equiv.) and(tert-butoxycarbonyl)glycylglycylglycine (1 equiv.) at 0° C. The mixtureis stirred at rt for 2 h. The mixture is quenched with H₂O, extractedwith DCM and washed with brine. The organic phase is dried andconcentrated. The residual is purified by flash column to givetert-butyl(2-((2-((2-((((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)carbamate.

Step 4: To a solution of tert-butyl(2-((2-((2-((((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)carbamatein DCM is added HCl (6M in H₂O) at rt under N₂. After stirring for 2 h,the mixture is quenched with NaHCO₃(aq) and extracted with EA. Thecombined organic layer is washed with brine, dried over anhydrousNa₂SO₄. The residue is concentrated and purified by chromatography onsilica gel (0-50% ethyl acetate in petroleum) to give2-amino-N-(2-((2-((((2R,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)amino)-2-oxoethyl)acetamide.

Step 5: To a solution of2-amino-N-(2-((2-((((2R,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)amino)-2-oxoethyl)acetamidein DMF is added DIPEA and(2S,4R)-1-(2-(3-acetyl-5-(2-fluoropyrimidin-5-yl)-1H-indazol-1-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide,then the mixture is stirred at reflux overnight. On consumption ofstarting material (LCMS monitoring), the reaction vessel is again cooledto room temperature. The mixture is diluted with ethyl acetate andwashed with water. The organic layer is dried over sodium sulfate,concentrated under reduced pressure to give a crude product, which ispurified by flash chromatography to give(2S,4R)-1-(2-(3-acetyl-5-(2-((2-((2-((2-((((2R,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-yl)methyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)-2-oxoethyl)amino)pyrimidin-5-yl)-1H-indazol-1-yl)acetyl)-N-(6-bromopyridin-2-yl)-4-fluoropyrrolidine-2-carboxamide(Compound 54).

Preparation of Compound 55:2-(2-(2-((2R,3S)-2-(4-(cyclopentylamino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carboxamido)-3-hydroxypropanamido)-3-(1H-indol-3-yl)propanamido)-N1-(1-((((2R,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-yl)methyl)amino)-4-(methylthio)-1-oxobutan-2-yl)succinamide

Step 1: To a solution of(2R,3S)-2-(4-(cyclopentylamino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carboxylicacid in EtOH is added DMAP and di-tert-butyl dicarbonate at 0° C. inportions. The mixture is vigorously stirred and allowed to warm up toroom temperature slowly overnight (16 h). The solvents are evaporatedunder vacuum and the residue is purified by column chromatography (SiO₂,solvent gradient: DCM to 1:9 MeOH/DCM) to give(2R,3S)-2-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carboxylicacid.

Step 2: To a solution of benzyl seryltryptophylasparaginylmethioninate(1 equiv.) in DMF is added HATU (1.5 equiv.), DIPEA (3 equiv.) and(2R,3S)-2-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carboxylicacid (1 equiv.) at 0° C. The mixture is stirred at rt for 2 h. Themixture is quenched with H₂O, extracted with DCM and washed with brine.The organic phase is dried and concentrated. The residual is purified byflash column to give benzyl((2R,3S)-2-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carbonyl)seryltryptophylasparaginylmethioninate.

Step 3: To a solution of benzyl((2R,3S)-2-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carbonyl)seryltryptophylasparaginylmethioninatein MeOH is added Pd/C at rt under a H₂ balloon. The reaction is stirredat rt for 1.5 h. The resulting mixture is filtered and concentrated invacuo. The crude product((2R,3S)-2-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carbonyl)seryltryptophylasparaginylmethionineis used to next step with no further purification.

Step 4: To a solution of((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methanamine(1 equiv.) in DMF is added HATU (1.5 equiv.), DIPEA (3 equiv.) and(((2R,3S)-2-(4-((tert-butoxycarbonyl)(cyclopentyl)amino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carbonyl)seryltryptophylasparaginylmethionine(1 equiv.) at 0° C. The mixture is stirred at rt for 2 h. The mixture isquenched with H₂O, extracted with DCM and washed with brine. The organicphase is dried and concentrated. The residual is purified by flashcolumn to give tert-butyl(4-((2R,3S)-3-((11-((1H-indol-3-yl)methyl)-8-(2-amino-2-oxoethyl)-5-((((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)carbamoyl)-15-hydroxy-7,10,13-trioxo-2-thia-6,9,12-triazapentadecan-14-yl)carbamoyl)-1-(2-fluoro-6-methylbenzoyl)piperidin-2-yl)phenyl)(cyclopentyl)carbamate.

Step 8: To a solution of tert-butyl(4-((2R,3S)-3-((11-((1H-indol-3-yl)methyl)-8-(2-amino-2-oxoethyl)-5-((((3aR,4R,7R,7aR)-2,2-dimethyl-7-(1-methyl-1H-pyrazol-4-yl)tetrahydro-4H-[1,3]dioxolo[4,5-c]pyran-4-yl)methyl)carbamoyl)-15-hydroxy-7,10,13-trioxo-2-thia-6,9,12-triazapentadecan-14-yl)carbamoyl)-1-(2-fluoro-6-methylbenzoyl)piperidin-2-yl)phenyl)(cyclopentyl)carbamatein THE is added HCl (2N in H₂O) at 0° C. The mixture is stirred at rtfor 2 h. The mixture is concentrated and purified by prep-HPLC (MethodA) to give2-(2-(2-((2R,3S)-2-(4-(cyclopentylamino)phenyl)-1-(2-fluoro-6-methylbenzoyl)piperidine-3-carboxamido)-3-hydroxypropanamido)-3-(1H-indol-3-yl)propanamido)-N1-(1-((((2R,3R,4R,5R)-3,4-dihydroxy-5-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-2-yl)methyl)amino)-4-(methylthio)-1-oxobutan-2-yl)succinamide(Compound 55).

Preparation of Compound 56:N-(11-(((2R,3R,4R,5S)-5-((4-chloro-3-(trifluoromethyl)pyridin-2-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((2-((3-((2-(((2R,3R,4R,5S)-5-((4-chloro-3-(trifluoromethyl)pyridin-2-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide

Step 1: To a solution of13-((2-carboxyethoxy)methyl)-1-((2,4-dinitrophenyl)amino)-11-oxo-3,6,9,15-tetraoxa-12-azaoctadecan-18-oicacid (1 equiv.) in DMF (2 mL) is added HATU (1.5 equiv.), DIPEA (3equiv.) and(2R,3R,4R,5S)-2-(((11-aminoundecyl)oxy)methyl)-5-((4-chloro-3-(trifluoromethyl)pyridin-2-yl)amino)tetrahydro-2H-pyran-3,4-diol(1 equiv.) at 0° C. The 10 mixture is stirred at rt for 2 h. The mixtureis quenched with H₂O, extracted with DCM and washed with brine. Theorganic phase is dried and concentrated. The residual is purified byflash column to giveN-(11-(((2R,3R,4R,5S)-5-((4-chloro-3-(trifluoromethyl)pyridin-2-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)-3-((2-((3-((2-(((2R,3R,4R,5S)-5-((4-chloro-3-(trifluoromethyl)pyridin-2-yl)amino)-3,4-dihydroxytetrahydro-2H-pyran-2-yl)methoxy)undecyl)amino)-3-oxopropoxy)methyl)-14-((2,4-dinitrophenyl)amino)-4-oxo-6,9,12-trioxa-3-azatetradecyl)oxy)propanamide(Compound 56).

Example 9. Non-limiting exemplary compounds of the present invention

TABLE 1 Degraders # Compound 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

TABLE 2 ASGPR Ligands of the Present Invention SPR Binding External CmpdCalculation: # Structure KD (uM) A90 

+++ A92 

+ A95 

++++ A96 

A97 

A98 

A99 

+ A100

++++ A101

A102

A103

+++ A104

A105

++++ A106

++++ A107

+ A108

++++ A109

++++ A110

++++ A111

A112

++++ A113

++++ A114

++++ A115

A116

++++ A117

+++ A118

++++ A119

++++ A120

+++ A121

+++ A127

++++ A128

+ A129

A134

A160

+ A162

+ A164

++ A169

A181

+++ A182

+ A183

+++ A184

++++ A187

++++ A191

+ A192

+ A193

+++ A196

++++ A209

+ A217

+ A283

+++ A284

+++ A285

++++ A286

++++ In the table above K_(D) values that are >= 1000 uM = +, <1000 uM =++, <500 uM = +++, and <100 uM = ++++

Example 10. Affinity of compounds to ASGPR measured using SurfacePlasmon Resonance (SPR)

The dissociation constants (KD) of compounds described herein to theASGP receptor are measured in SPR experiments using a Biacore 8Kinstrument (GE Healthcare) at 25° C. Recombinant ASGPR protein is firstbiotinylated using Maleimide-PEG2-biotin reagent (Pierce, 19-fold molarexcess) in phosphate-buffered saline (PBS) solution overnight at 4° C.Excess amount of biotin in the reaction mixture is removed by Zebadesalting columns (Thermo). Biotinylation is confirmed by massspectroscopic analysis of ASGPR. Biotinylated ASGPR is then immobilizedon SA sensor chips (GE Healthcare) with an immobilization level rangingfrom 1500-3000 resonance units (RU). The running buffer is 50 mM Tris,pH7.5, 150 mM NaCl, 50 mM CaCl₂), 0.01% P20, 3% DMSO. The concentrationof compounds sometimes vary from 2 mM to 50 μM depending on KD values.The compounds are diluted 3 folds with total 8 concentration points.Solutions containing serially diluted compounds are injected at a flowrate of 50 μL/min for 60 sec followed by a 180 sec dissociation phasefor each concentration. Data is processed using the analysis software inBiacore 8K to perform background subtraction, double referencing andsolvent correction. Values of affinity expressed as the dissociationconstants (KD) were determined by fitting the steady state bindingresponses (RUss) as a function of the concentration ([Compound]) usingthe following equation: RUss=RUmax/(KD+[Compound]), where RUmax is thecalculated maximal response.

TABLE 3 Biological Data for Extracellular Protein Degrading CompoundsSPR Binding External SPR Binding External Calculation: KD (uM)Calculation: KD (uM) Compound # ASGPR Additional Targets Compound 20++++ ++++ hIgA1-FL Compound 21 ++++ ++++ hIgA1-FL Compound 22 ++++ ++++hIgA1-FL Compound 23 ++++ ++++ hIgA1-Fc Compound 24 ++++ ++++ hIgA1-FcCompound 25 ++++ ++++ hIgA1-Fc Compound 26 ++++ Compound 27 ++++hIgA1-Fc In the table above KD values that are >= 1000 uM = +, <1000 uM= ++, <500 uM = +++, and <100 uM = ++++

TABLE 4 Biological Data for ASGPR Ligands ASGPR_PR Binding Compound IDStructure K_(D) (μM) A127-2c

 >200 A131

+++ A136

+++ A146

++++ A147

++++ A148

>20 A149

>20 A153

+++ A157

>20 A163

 >200 A170

 >200 A174

++++ A180

++++ A186

+ A188

+++ A212

+++ A213

+++ A218

+ A221-2

>200 A222

+++ A223

++++ A287

++++ A288

++++ A289

++++ A290

++++ A291

++++ A292

++++ A293

++++ A294

++++ A295

++++ A296

++++ A298

++++ A299

++++ A300

++++ A301

++++ A302

++++ A304

++++ A305

++++ A305

>20 A306

++++ A307

++++ A308

>20 A309

++++ A310

>20 A311

>20 A312

++++ A313

>20 A314

>20 A315

++++ A316

>20 A317

++++ A318

++++ A319

++++ A320

++++ A321

++++ A322

>20 A323

++++ A324

++++ A325

++++ A326

++++ A327

++++ A328

>20 A330

++++ A331

++++ A332

++++ A333

>10 A334

++++ A335

++++ A336

++++ A338

++++ A339

++++ A340

++++ A341

++++ A342

++++ A344

++++ A345

++++ A346

++++ A348

>20 A349

++++ A350

>20 A351

>20 A352

>20 A353

A354

++++ A355

++++ A356

>20 A358

++++ A359

++++ A360

++++ A361

>20 A362

++++ A363

++++ A364

++++ A365

>20 A366

++++ A367

++++ A368

>20 A369

>20 A370

>10 A371

++++ A372

>20 A373

++++ A374

>20 A377

++++ A378

++++ A379

++++ A380

++++ A381

++++ A382

++++ A383

++++ A384

++++ A385

 >200 A386

>20 A387

>20 A388

>20 A389

>20 A390

 >200 A392

+++ A393

+++ A394

 >200 A395

 >200 A397

+++ A399

 >200 A400

 >200 A401

 >200 A402

 >200 A403

>20 A404

++++ A405

++++ A406

++++ A425

+ A426

++++ A427

++++ A91 

++++ In the table above KD values that are >= 1000 uM = +, <1000 uM =++, <500 uM = +++, and <100 uM = ++++

TABLE 5 Biological Data for ASGPR Ligands with Linker ASGPR_SPR BindingCompound ID K_(D) (μM) Structure Compound 29

Compound 30 ++++

Compound 31 ++++

Compound 32

Compound 33 ++++

Compound 34

A413 ++++

Compound 35 ++++

Compound 36

Compound 37

Compound 38

Compound 39

Compound 56

A420

A421

A423

Compound 48

Compound 49

Compound 50

A417 ++++

A424

A422 ++++

In the table above KD values that are >= 1000 uM = +, <1000 uM = ++,<500 uM = +++, and <100 uM = ++++

Example 11: Cellular Degradation of an Extracellular Protein byASGPR-binding Extracellular Protein Targeting Ligand Degrader Compoundsas Described Herein

The selected Extracellular Target Protein (typically 80 uM to 1 mM) isadded to cell culture media in the presence or absence of thebifunctional degrader as described herein. The assay is performed withhuman hepatic cell line HepG2 in 96-well plates. HepG2 cells is culturedto 70-80% confluency in RPMI media (ThermoFisher/Gibco). The cells arewashed two times with PBS solution and then treated with serum-freemedia containing human IgA or other target proteins. The bifunctionaldegrader is then added to the cell culture media with the topconcentration of 20 μM in a 2-fold dilution series. The cells are thenincubated for 24h at 37° C. An aliquot of the supernatant is thenremoved, diluted (10 to 100-fold dilution) and the concentration of thetarget protein analyzed with a sandwiched ELISA assay using a commercialkit (MyBioSource or equivalent) in a 96-well plate. Dose-dependentdepletion of the target protein is analyzed by GraphPad Prism softwareand the data were fitted to a sigmoidal curve to obtain IC50 values.

Example 12. Cellular Degradation of IgA by Bifunctional MoleculesContaining ASGPR and Target Protein-Binding Moieties

Human IgA (Sigma) protein (80 uM to 1 mM) was added to the cell culturemedia in the presence or absence of Compound A below that includes IgAbinding peptide OPT-3. The studies were performed with human hepaticcell line HepG2 in 96-well plates. HepG2 cells were cultured to 70-80%confluency in RPMI media (ThermoFisher/Gibco). The cells were washed 2times with PBS solution and were then treated with serum-free mediacontaining human IgA or other target proteins. The bifunctionalcompounds were then added to the cell culture media with the topconcentration of 20 μM in a 2-fold dilution series. The cells were thenincubated for 24h at 37° C. An aliquot of the supernatant was thenremoved, diluted (10 to 100-fold dilution) and the concentration of thetarget protein was analyzed by a sandwiched ELISA assay using acommercial kit (MyBioSource or equivalent) in a 96-well plate.Dose-dependent depletion of the target protein was analyzed by GraphPadPrism software and the data were fitted to a sigmoidal curve to obtainIC50 values.

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference.

Although the foregoing invention has been described in some detail byway of illustration and example for the purpose of clarity ofunderstanding, it will be readily apparent to one of ordinary skill inthe art in light of the teaching of this invention that certain changesor modifications may be made thereto without departing from the spiritor scope of the invention as defined in the appended claims.Additionally, those skilled in the art will recognize, or be able toascertain using no more than routine experimentation, many equivalentsto the specific embodiments and methods described herein. Suchequivalents are intended to be encompassed by the scope of the presentapplication.

We claim:
 1. A compound of formula:

or a pharmaceutically acceptable salt thereof; PGP-ll28,C₁ wherein: R¹is selected from the group consisting of hydrogen, alkyl, alkenyl, andhaloalkyl; R² is selected from the group consisting of —NR⁶-heteroaryland—NR⁶-aryl, each of which R² is optionally substituted with 1, 2, 3,or 4 substituents independently selected from the group consisting ofalkyl, alkenyl, haloalkyl, —OR⁶, F, Cl, Br, I, —NR⁶R⁷, heteroalkyl,cyano, nitro, and C(O)R³; R³ at each occurrence is independentlyselected from the group consisting of hydrogen, alkyl, heteroalkyl,haloalkyl, arylalkyl, alkenyl, aryl, heteroaryl, heterocycle, —OR⁸,and—NR⁸R⁹; R⁶ and R⁷ are independently selected at each occurrence fromthe group consisting of hydrogen, alkyl, arylalkyl, alkenyl, aryl,haloalkyl, heteroaryl, heterocycle, and C(O)R³; R⁸ and R⁹ areindependently selected at each occurrence from the group consisting ofhydrogen, alkyl, arylalkyl, alkenyl, aryl, heteroaryl, and heterocycle;Linker^(A) is bond; Linker^(B) is

R¹¹, R¹², R¹³, R¹⁴, R¹⁵, R¹⁶, R¹⁷, R¹”, R¹⁹, and R²⁰ are independentlyat each occurrence selected from the group consisting of a bond, alkyl,—C(O)—, —C(O)O—, —OC(O)—, —SO₂—, —S(O)—, —C(S)—, —C(O)NR⁶—, —NR⁶C(O)—,—O—, —S—, —NR⁶—, —C(R²¹R²¹)—, —P(O)(R³)O—, —P(O)(R³)—, alkenyl, alkynyl,haloalkyl, alkoxy, aryl, heterocycle, heteroaryl,—CH₂CH₂—[O—(CH₂)₂]_(n)—O—, —CH₂CH₂—[O—(CH₂)₂]_(n)—NR⁶—,—CH₂CH₂—[O—(CH₂)₂]_(n)—, —[—(CH₂)₂—O—]_(n)—, —[O—(CH₂)₂]_(n)—,—[O—CH(CH₃)C(O)]_(n)—, —[C(O)—CH(CH₃)—O],—, —[O—CH₂C(O)]_(n)—,and—[C(O)—CH₂—O]_(n)—, each of which is optionally substituted with 1,2, 3, or 4 substituents independently selected from R²¹; n isindependently selected at each instance from 0, 1, 2, 3, 4, 5, 6, 7, 8,9, or 10; R²¹ is independently at each occurrence selected from thegroup consisting of hydrogen, alkyl, alkenyl, F, Cl, Br, I, hydroxyl,alkoxy, azide, amino, cyano, —NR⁶R⁷, —NR⁸SO₂R³, —NR'S(O)R³, haloalkyl,aryl, heteroaryl, and heterocycle; Linker^(C) is selected from:

R²² is selected from the group consisting of alkyl, —C(O)N—, —NC(O)—,—N—, —C(R²¹)—, —P(O)—, alkenyl, haloalkyl, aryl, heterocycle, andheteroaryl, each of which is optionally substituted with 1, 2, 3, or 4substituents independently selected from R²¹; and Extracellular ProteinTargeting Ligand is a chemical moiety that binds to the targetedextracellular protein that creates or exacerbates a disease.
 2. Thecompound of claim 1, wherein R² is —NR⁶-heteroaryl optionallysubstituted with 1, 2, or 3 substituents independently selected from thegroup consisting of alkyl, alkenyl, haloalkyl, —OR⁶, F, Cl, Br, —NR⁶R⁷,heteroalkyl, cyano, nitro, and C(O)R³.
 3. The compound of claim 2,wherein R⁶ is hydrogen.
 4. The compound of claim 3, wherein R¹ ishydrogen.
 5. The compound of claim 1, wherein R² is —NR⁶-heteroarylsubstituted with 1 or 2, substituents independently selected from thegroup consisting of alkyl, haloalkyl, —OR⁶, F, C₁, and—NR⁶R⁷.
 6. Thecompound of claim 5, wherein R⁶ is hydrogen.
 7. The compound of claim 6,wherein heteroaryl is selected from the group consisting of


8. The compound of claim 7, wherein the 1 or 2 substituents areindependently selected from the group consisting of haloalkyl andhalogen.
 9. The compound of claim 8, wherein halogen is fluorine orchloride.
 10. The compound of claim 6, wherein heteroaryl is selectedfrom the group consisting of


11. The compound of claim 10, wherein the 1 or 2 substituents areindependently selected from the group consisting of haloalkyl andhalogen.
 12. The compound of claim 1, wherein R² is —NR⁶-aryl.
 13. Thecompound of claim 1 Formula:

or a pharmaceutically acceptable salt thereof.
 14. The compound of claim13, wherein R² is —NR⁶-heteroaryl optionally substituted with 1, 2, or 3substituents independently selected from the group consisting of alkyl,alkenyl, haloalkyl, —OR⁶, F, Cl, Br, —NR⁶R⁷, heteroalkyl, cyano, nitro,and C(O)R³.
 15. The compound of claim 14, wherein Linker^(B) is selectedfrom the group consisting of


16. The compound of claim 14, wherein Linker^(B) is selected from thegroup consisting of


17. The compound of claim 1 of Formula:

or a pharmaceutically acceptable salt thereof.
 18. The compound of claim17, wherein R² is —NR⁶-heteroaryl optionally substituted with 1, 2, or 3substituents independently selected from the group consisting of alkyl,alkenyl, haloalkyl, —OR⁶, F, Cl, Br, I, —NR⁶R⁷, heteroalkyl, cyano,nitro, and C(O)R³.
 19. The compound of claim 18, wherein Linker^(C) isselected from:


20. The compound of claim 18, wherein Linker^(C) is selected from:


21. The compound of claim 1, wherein the Extracellular Protein TargetingLigand targets an immunoglobin.
 22. The compound of claim 1, wherein theExtracellular Protein Targeting Ligand targets IgA.
 23. The compound ofclaim 1, wherein the Extracellular Protein Targeting Ligand targets IgG.24. The compound of claim 1, wherein the Extracellular Protein TargetingLigand targets IgE.
 25. The compound of claim 1, wherein theExtracellular Protein Targeting Ligand targets TNF-α.
 26. The compoundof claim 1, wherein the Extracellular Protein Targeting Ligand targetsIL-1b, IL-2, or IL-6.
 27. The compound of claim 1, wherein theExtracellular Protein Targeting Ligand targets IFN-γ or VEGF.
 28. Thecompound of claim 1, wherein the Extracellular Protein Targeting Ligandtargets TGF-b1.
 29. The compound of claim 1, wherein the ExtracellularProtein Targeting Ligand targets PCSK-9.
 30. A pharmaceuticalcomposition comprising a compound of claim 1 or a pharmaceuticallyacceptable salt thereof and a pharmaceutically acceptable carrier.